Professional Documents
Culture Documents
Author manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
Author Manuscript
Abstract
The extracellular domain of several membrane-anchored proteins is released from the cell surface
as soluble proteins through a regulated proteolytic mechanism called ectodomain shedding. Cells
use ectodomain shedding to actively regulate the expression and function of surface molecules,
and modulate a wide variety of cellular and physiological processes. Ectodomain shedding rapidly
converts membrane-associated proteins into soluble effectors and, at the same time, rapidly
reduces the level of cell surface expression. For some proteins, ectodomain shedding is also a
prerequisite for intramembrane proteolysis, which liberates the cytoplasmic domain of the affected
molecule and associated signaling factors to regulate transcription. Ectodomain shedding is a
process that is highly regulated by specific agonists, antagonists, and intracellular signaling
Author Manuscript
pathways. Moreover, only about 2% of cell surface proteins are released from the surface by
ectodomain shedding, indicating that cells selectively shed their protein ectodomains. This review
will describe the molecular and cellular mechanisms of ectodomain shedding, and discuss its
major functions in lung development and disease.
Keywords
ectodomain shedding; lung injury; inflammation; infection; host defense
factors, cell adhesion molecules, enzymes, and many more (Table 1). There are several
common mechanistic features in the ectodomain shedding of cell surface molecules (Fig. 1).
Most show little basal shedding, but are dramatically induced upon cellular activation.
Phorbol esters induce shedding of the majority of protein ectodomains, suggesting that
protein kinase C (PKC) is one of the key regulators. Indeed, chemically mutagenized CHO
cells that are defective in PMA-induced TGFα shedding are also defective in phorbol 12-
myristate 13-acetate (PMA)-induced shedding of several other transmembrane proteins,
*
Correspondence to: Pyong Woo Park, Ph.D., Children’s Hospital, Harvard Medical School, 320 Longwood Avenue, Enders-144,
Boston, MA 02115. Fax: (617)730-0240. pyong.park@childrens.harvard.edu.
HAYASHIDA et al. Page 2
such as TNFα, amyloid precursor protein (APP), and L-selectin (Arribas and Massague,
Author Manuscript
1995; Arribas et al., 1996). Furthermore, inhibitors of protein tyrosine kinases (PTKs;
Fitzgerald et al., 2000; Gutwein et al., 2000) and MAP kinases inhibit shedding (Fan and
Derynck, 1999; Fitzgerald et al., 2000), whereas agonists of G-protein coupled receptors
(GPCRs), calcium ionophores, and ceramide induce shedding of a number of molecules
(Porteu and Nathan, 1990; Madge et al., 1999; Fitzgerald et al., 2000; Lemjabbar and
Basbaum, 2002; Matthews et al., 2003; Mortier et al., 2004). These data illustrate that
several intracellular signaling pathways regulate ectodomain shedding and, consistent with
these observations, agonists that activate these signaling pathways, such as growth factors,
cytokines, and bacterial toxins, have been shown to enhance shedding (Subramanian et al.,
1997; Jones et al., 1999; Yabkowitz et al., 1999; Fitzgerald et al., 2000; Park et al., 2000,
2004; Chen et al., 2007). In addition, peptide hydroxamate inhibitors of metalloproteinases
inhibit the shedding of most cell surface proteins, with the exception of
Author Manuscript
(Fitzgerald et al., 2000), calcium ionophores (Dethlefsen et al., 1998; Reiss et al., 2005), and
bacterial toxins (Walev et al., 1996; Park et al., 2000, 2004; Chen et al., 2007). Some of
these globally activate shedding of many cell surface proteins, whereas others specifically
stimulate shedding of certain protein ectodomains. Rapid induction of shedding by phorbol
esters is one of the hallmark features of ectodomain shedding and, accordingly, several PKC
isozymes regulate shedding. For example, PKCε is required for TNFα shedding (Wheeler et
al., 2003). PKCδ regulates heparin binding (HB) epidermal growth factor (EGF)-like
shedding (Izumi et al., 1998), whereas PKCδ and PKCη are involved in IL-6 receptor
(IL-6R) shedding (Thabard et al., 2001). These findings indicate that different substrates
require distinct PKC isozymes for its shedding regulation, but the underlying basis of this
specificity is not understood.
Several studies indicate that PTKs are also positive regulators of ectodomain shedding
Author Manuscript
(Subramanian et al., 1997; Fitzgerald et al., 2000; Gutwein et al., 2000; Phong et al., 2003).
Pervanadate, a general and potent inhibitor of protein tyrosine phosphatases, enhances the
shedding of the adhesion molecule L1, APP, syndecan-1 and -4, and angiotensin converting
enzyme (ACE), among others, and PTK inhibitors inhibit the shedding of these surface
molecules (Subramanian et al., 1997; Fitzgerald et al., 2000; Gutwein et al., 2000; Park et
al., 2000; Phong et al., 2003). Precisely how kinases modulate shedding has yet to be clearly
defined, but current data suggest that both PTKs and PKCs do not enhance shedding by
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 3
phosphorylating the substrate. For example, pervanadate-induced ACE shedding does not
Author Manuscript
require the cytoplasmic domain of ACE (Santhamma et al., 2004). Similarly, deletion of the
cytoplasmic tails of IL-6R (Mullberg et al., 1994) and TNFα receptor II (TNFRII) (Crowe et
al., 1993) does not affect shedding of their ectodomains by PMA, despite the fact that the
cytoplasmic domains of these cytokine receptors are phosphorylated by PMA stimulation.
Antibody crosslinking induces the shedding of L-selectin by sequestering L-selectin to a
lipid raft membrane compartment where L-selectin is highly Tyr phosphorylated (Phong et
al., 2003), but there is no evidence that Tyr phosphorylation of the L1 cytoplasmic domain is
required for shedding. Consistent with these data, the cytoplasmic tail of L1 is not Tyr
phosphorylated in PMA-induced shedding (Gutwein et al., 2000), and Tyr residues of the L1
cytoplasmic domain are dispensable in hepatocyte growth factor (HGF)-induced shedding
(Heiz et al., 2004). Shedding of APP induced by pervanadate and PMA stimulation is
blocked by PTK inhibitors, but the cytoplasmic domain of APP is not Tyr phosphorylated by
Author Manuscript
pervanadate or PMA stimulation (Slack et al., 1995). Further, PMA and several other
inducers of HB-EGF shedding induce Ser phosphorylation of the HB-EGF cytoplasmic
domain, but mutation of the phosphorylated Ser residues has no effect on HB-EGF shedding
(Wang et al., 2006). Collectively, these data indicate that PKCs and PTKs phosphorylate
components other than the substrate in enhancing ectodomain shedding.
The cytoplasmic domain of substrates, however, can affect ectodomain shedding through
interactions with intracellular modifiers. Calmodulin binds to the cytoplasmic domain of L-
selectin and ACE in a constitutive manner, and its dissociation induced by calmodulin
kinase inhibitors or PMA activates shedding, indicating that calmodulin is a negative
regulator of L-selectin and ACE shedding (Kahn et al., 1998; Matala et al., 2001). In fact,
calmodulin kinase inhibitors and calcium ionophores also induce the shedding of CD44,
EGF, betacellulin, N-cadherin, and IL-6R (Nagano et al., 2004; Reiss et al., 2005; Sanderson
Author Manuscript
et al., 2005). These data suggest that regulation of shedding by calmodulin and calcium is a
common mechanism. The APP homolog APLP1 binds to APP and interferes with
endocytosis of APP through a conserved NPTY motif in the cytoplasmic domain and
increases APP shedding (Neumann et al., 2006). The carboxyl terminus Val in the
cytoplasmic domain of TGFα is required for its shedding induced by various stimuli
(Bosenberg et al., 1992), suggesting that structural features of the TGFα cytoplasmic tail is
essential for its interaction with intracellular regulators of shedding. Available data also
implicate actin-binding proteins of the ERM family in the regulation of shedding. Ezrin is
bound to the cytoplasmic tail of L-selectin in both resting and PMA-stimulated lymphocytes,
but only moesin binding to the L-selectin cytoplasmic tail is associated with the activation of
shedding (Ivetic et al., 2002). Importantly, mutation of the Ezrin/Radixin/Moesin (ERM)
binding site interferes with L-selectin shedding, suggesting that ERM interactions are
Author Manuscript
In addition to PKC and PTK signaling, other signaling pathways involving ATP
(Hubschmann et al., 2005; Sengstake et al., 2006), MAP kinases (Fan and Derynck, 1999;
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 4
Fitzgerald et al., 2000), and GPCRs (Prenzel et al., 1999; Fitzgerald et al., 2000) have been
Author Manuscript
Sheddases
The first clue to identify the sheddase was provided by the finding that peptide hydroxamate
inhibitors of metalloproteinases block the shedding of membrane bound TNFα (Mohler et
al., 1994). Subsequently, shedding of the majority of proteins was also found to be inhibited
by peptide hydroxamates, and TNFα converting enzyme (TACE) was identified as the
TNFα sheddase in 1997 (Black et al., 1997; Moss et al., 1997). TACE is a member of the
disintegrin and metalloproteinase (ADAM) family (Schlondorff and Blobel, 1999), and is
also referred to as ADAM17. TACE/ADAM17 is also a sheddase for other surface
molecules, such as TNFRs (Peschon et al., 1998), L-selectin (Peschon et al., 1998), vascular
cell adhesion molecule-1 (VCAM-1; Garton et al., 2003), fractalkine (Garton et al., 2001),
Author Manuscript
ErbB-4 (Rio et al., 2000), colony stimulating factor-1 (CSF-1) (Horiuchi et al., 2007b), APP
(Buxbaum et al., 1998; Huovila et al., 2005), and the EGF family ligands TGFα (Peschon et
al., 1998), HB-EGF (Sunnarborg et al., 2002; Jackson et al., 2003), amphiregulin
(Sunnarborg et al., 2002; Sternlicht et al., 2005), and epiregulin (Sahin et al., 2004).
Although TACE/ADAM17 can shed many cell surface proteins, it is not the only sheddase
and several proteins that are not shed by TACE/ADAM17 (e.g., ACE, TRANCE,
syndecan-1) (Sadhukhan et al., 1999; Fitzgerald et al., 2000; Schlondorff et al., 2001).
ADAM9 and ADAM12 shed HB-EGF (Izumi et al., 1998; Asakura et al., 2002), ADAM10
sheds E-cadherin (Maretzky et al., 2005), N-cadherin (Reiss et al., 2005), HB-EGF
(Lemjabbar and Basbaum, 2002), betacellulin (Horiuchi et al., 2007a), Notch (Pan and
Rubin, 1997), L1 (Mechtersheimer et al., 2001), APP (Huovila et al., 2005) and CXCL16
(Abel et al., 2004), and ADAM8, ADAM15, and ADAM28 shed CD23, a low affinity IgE
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 5
Bredow et al., 1997), and syndecan-1 (Li et al., 2002), and MMP-9 sheds E-cadherin
Author Manuscript
(Symowicz et al., 2007), ICAM-1 (Fiore et al., 2002), c-kit ligand (Heissig et al., 2002), and
syndecan-1 and -4 (Brule et al., 2006). The transmembrane MMP, MMP-14, can mediate the
shedding of TRANCE (Schlondorff et al., 2001), CD44, and syndecan-1. In addition, many
bacterial pathogens, including Pseudomonas aeruginosa, E. coli, Bacteroides fragilis,
Staphylococcus aureus, Streptococcus pyogenes (Group A Streptococcus, GAS), Listeria
monocytogenes, Bacillus anthracis, and Streptococcus pneumoniae, express
metalloproteinases that shed various inflammatory factors from the host cell surface
(Vollmer et al., 1996; Lathem et al., 2003; Grys et al., 2005; Chung et al., 2006; Chen et al.,
2007; Leduc et al., 2007; Wu et al., 2007). Thus, many metalloproteinases possess the
capacity to function as sheddases. However, ectodomain shedding is likely specific in vivo
because the expression of the substrate, sheddase, and extracellular and intracellular
regulatory factors are tightly controlled in a spatial and temporal manner.
Author Manuscript
acids is not shed efficiently (Elenius et al., 1997). Further, a mutant construct of APP with
deletion of the cleavage site is still cleaved at a different -P1- - sequence at exactly the
same distance from the membrane as ™-type APP (Maruyama et al., 1991). However, a 14-
amino acid sequence of the juxtamembrane domain of APP or TGFα is sufficient to confer
shedding susceptibility to betaglycan (Arribas et al., 1997), whereas an 11 amino acid
deletion within the juxtamembrane region of APP inhibited its shedding by α secretase
(Sisodia et al., 1990). Thus, although the cleavage site is predominantly specified by the
distance from the plasma membrane and structure of the cleavage site region, these latter
findings suggest that there is some degree of sequence specificity in the shedding of certain
substrates.
ADAMs and MMPs are produced as zymogens, and proteolytic removal of the prodomain
activates these metalloproteinases. ADAMs typically have a cleavage site in their prodomain
for the serine endopeptidases, furin and furin-like proprotein convertase (PC), and several
studies have shown that ADAM zymogens are activated through cleavage of the prodomain
by furin-like PCs in the trans-Golgi/endosomal compartment (Lum et al., 1998; Loechel et
al., 1999; Roghani et al., 1999). Although proteolytic processing by furins is generally
constitutive and occurs in intracellular compartments, several studies have shown that furin
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 6
can process proteins in a specific cellular compartment formed by the fusion of endocytic
Author Manuscript
inhibitory effect of the prodomain on the enzymatic activity (Zhang et al., 2001).
Some studies have also shown that intracellular trafficking of sheddases influences
ectodomain shedding. For example, PMA treatment induces the translocation of ADAM12
to the cell surface in a PKCε-dependent manner (Sundberg et al., 2004). Another study
showed that in polarized epithelial cells, ADAM10 is sorted exclusively to the basolateral
compartment and this depends on specific Pro residues in the Src homology 3 binding
domain (SH3) in the cytoplasmic domain of ADAM10 (Wild-Bode et al., 2006).
Interestingly, this SH3 motif is also required for ADAM10-mediated shedding of E-cadherin
(Wild-Bode et al., 2006). In addition, ethyl-methane sulfonate-induced CHO mutant cells
defective in TACE/ADAM17-dependent ectodomain shedding have impaired trafficking of
TACE/ADAM17 from the ER, which interferes with the activation of TACE/ADAM17 by
furins (Borroto et al., 2003).
Author Manuscript
Cells also regulate shedding by targeting the substrate to specific cellular compartments.
Cholesterol depletion from cells, which displaces substrates from lipid raft microdomains,
induces the shedding of CD30 by TACE/ADAM17 and IL-6 receptor by ADAM10 and
TACE/ADAM17 (Matthews et al., 2003; von Tresckow et al., 2004), and antibody-induced
clustering induces shedding of CD30 (Hansen et al., 2000), CD44 (Shi et al., 2001), and L-
selectin (Phong et al., 2003). Protein–protein interactions between the substrate and
modifying proteins also influence shedding. Substrate interactions with signaling molecules,
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 7
In tissue injury and inflammation, many immunomodulators are released from the cell
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 8
soluble TNFα, EGF, or TGFα (Zhao et al., 2001a, b), but the expression level of TGFα is
not affected in taceΔZn/ΔZn mice, suggesting that the pathologies are strictly caused by the
absence of TACE/ADAM17-mediated shedding of these cytokines and growth factors.
Moreover, the phenotypes of taceΔZn/ΔZn mice are similar to those of the TGFα null mice.
Strong TACE/ADAM17 expression is detected as early as E12, primarily on epithelial cell
surfaces. At E16.5, TACE/ADAM17 is ubiquitously expressed on both lung epithelial and
mesenchymal cells, but at E18, the expression in mesenchymal cells is significantly reduced.
Collectively, these observations suggest that TACE/ADAM17-mediated ectodomain
shedding of several growth factors and cytokines is essential for normal epithelial cell
proliferation and differentiation, branching morphogenesis, and vasculogenesis in the lung.
Ectodomain shedding of several cell surface molecules has been linked to the pathogenesis
of inflammatory lung diseases. In bleomycin-induced acute lung injury in mice, shedding of
syndecan-1 by MMP-7 generates a CXC chemokine gradient that guides the transepithelial
migration of neutrophils into the alveolar compartment (Li et al., 2002). In this mechanism,
injury caused by bleomycin induces the expression of the CXC chemokine KC (CXCL1,
mouse functional homolog of human IL-8) and MMP-7. Newly synthesized KC binds to the
heparan sulfate glycosaminoglycans of cell surface syndecan-1, and shedding of the
syndecan-1 ectodomain-KC complex by MMP-7 into the alveolar space generates a CXC
chemokine gradient across the alveolar epithelial border. The observations that both
syndecan-1 and MMP-7 null mice show increased accumulation of neutrophils in the
perivascular interstitial space, and significantly reduced neutrophils in the alveolar
compartment underscore the importance of this mechanism in coordinating inflammation
Author Manuscript
and confining inflammation to specific sites of tissue injury. Interestingly, though MMP-7
null mice are protected, syndecan-1 null mice show enhanced lung damage and lethality in
bleomycin-induced acute lung injury. These data suggest that syndecan-1 ectodomains have
additional protective functions in the progression of bleomycin-induced acute lung injury
downstream of its shedding by MMP-7.
Shedding of syndecan-1 also plays a protective role in allergic lung inflammation (Xu et al.,
2005). Intranasal challenge of mice with Aspergillus spp. culture filtrates induces features of
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 9
allergic lung inflammation, such as increased airway hyperresponsiveness, and Th2 cell
Author Manuscript
homing to the lung. Airway allergen challenge activates syndecan-1 shedding, and
syndecan-1 ectodomains bind to and inhibit the CC chemokines, CCL7, CCL11, and
CCL17, to recruit Th2 cells into the lung. Consistent with this mechanism, syndecan-1 null
mice show exaggerated airway hyperresponsiveness, glycoprotein hypersecretion,
eosinophilia, and Th2 responses in the lung relative to wild-type mice, and airway
administration of purified syndecan-1 ectodomains or heparan sulfate rescues allergen-
challenged syndecan-1 null mice from these inflammatory phenotypes. These data suggest
that syndecan-1 suppresses allergic lung inflammation by directly inhibiting CC chemokine-
induced Th2 cell homing in a heparan sulfate-dependent manner.
Several studies have shown that bacterial pathogens subvert ectodomain shedding to
enhance their virulence in the lung. For example, lipoteichoic acid released from the cell
wall of S. aureus stimulates ADAM-10 mediated shedding of HB-EGF ectodomains, which
Author Manuscript
activate the EGF receptor to induce mucin overexpression and subsequent airflow
obstruction in chronic lung infections (Lemjabbar and Basbaum, 2002). Streptolysin O, a
virulence factor toxin secreted by GAS, stimulates the ectodomain shedding of L-selectin
(Walev et al., 2000), IL-6R (Walev et al., 1996) and CD14 (Walev et al., 1996), suggesting
that streptolysin O-induced shedding modulates host defenses against GAS infections. P.
aeruginosa enhances syndecan-1 shedding by activating the endogenous shedding
mechanism through its virulence factor LasA (Park et al., 2000), and syndecan-1
ectodomains promote P. aeruginosa lung pathogenesis by inhibiting several host defense
mechanisms and dysregulating the host inflammatory response to infection (Park et al.,
2001). The physiological significance of this mechanism is underscored by the observations
that syndecan-1 null mice resist intranasal P. aeruginosa lung infection compared to wild-
type mice, and intranasal administration of syndecan-1 ectodomain or heparan sulfate
Author Manuscript
restores bacterial virulence in the lung (Park et al., 2001). Interestingly, S. aureus and S.
pneumoniae, but not other Gram-positive and Gram-negative bacteria, also enhance
syndecan-1 shedding (Park et al., 2004; Chen et al., 2007) and syndecan-1 null mice resist
lung infection caused by these major lung bacterial pathogens. These findings suggest that
bacterial pathogens with the capacity to enhance syndecan-1 ectodomain shedding do so to
promote their pathogenesis by dysregulating the host inflammatory response to infections.
selectively used in the host defense against Gram-negative pneumonia. CD44 null mice also
show exaggerated inflammation following bleomycin-induced lung injury, characterized by
impaired clearance of apoptotic neutrophils, persistent accumulation of hyaluronan
fragments at the site of tissue injury, and impaired activation of TGFβ (Teder et al., 2002).
Thus, in contrast to the shedding of syndecan-1, CD44 shedding apparently plays a
protective role in both infectious and non-infectious lung injury and inflammation.
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 10
CONCLUSIONS
Author Manuscript
A wide variety of cell surface proteins are proteolytically cleaved to release their
ectodomains into the extracellular milieu. Ectodomain shedding is an important
posttranslational modification that adds diversity to the function of cell surface molecules.
Ectodomain shedding rapidly downregulates the expression of cell surface proteins, and
liberates biologically active soluble ectodomains that can function in an autocrine or
paracrine manner. Once released, protein ectodomains exhibit functions similar to or distinct
from its cell surface counterpart. Furthermore, recent studies have shown that some
pathogens subvert ectodomain shedding to promote their infection. Regulated shedding is
typically a mechanism that modulates cellular processes, such as adhesion, migration and
proliferation, and assures the correct functioning of development and inflammation. Thus,
dysregulation or lack of shedding results in diverse pathologies such as inflammatory lung
injury, infection, cancer, and Alzheimer’s disease.
Author Manuscript
Studies during the last two decades have revealed many mechanistic features of ectodomain
shedding. Some are common, whereas others are specific to certain substrates. In general,
ectodomain shedding is dependent on PKC and metalloproteinase activities. However, PKC
isozymes selectively regulate the shedding of certain protein ectodomains, and numerous
metalloproteinases can function as sheddases, though available data suggest that TACE/
ADAM17 and ADAM10 are the primary sheddases for most ectodomains. Furthermore,
several other signaling factors and modifiers, such as PTKs, MAP kinases, calmodulin, BiP
and ARTS-1, are selectively used to regulate the shedding of certain cell surface molecules.
In addition, cellular processes affecting the recruitment, activation, or polarized secretion of
sheddases or substrates can also influence shedding, adding to the complexity of the
shedding mechanism. Future studies directed at defining how these regulatory factors and
Author Manuscript
processes coordinate ectodomain shedding should provide mechanistic insights into how
cells utilize ectodomain shedding to modulate diverse pathophysiological processes.
Acknowledgments
Grant sponsor: National Institutes of Health; Grant numbers: HL69050, HL73725, and HL81474; Grant sponsor:
Mizutani Foundation; Grant number: 070028.
LITERATURE CITED
Abel S, Hundhausen C, Mentlein R, Schulte A, Berkhout TA, Broadway N, Hartmann D, Sedlacek R,
Dietrich S, Muetze B, Schuster B, Kallen KJ, Saftig P, Rose-John S, Ludwig A. The transmembrane
CXC-chemokine ligand 16 is induced by IFN-gamma and TNF-alpha and shed by the activity of the
disintegrin-like metalloproteinase ADAM10. J Immunol. 2004; 172:6362–6372. [PubMed:
Author Manuscript
15128827]
Ahmed Z, Mazibrada G, Seabright RJ, Dent RG, Berry M, Logan A. TACE-induced cleavage of NgR
and p75NTR in dorsal root ganglion cultures disinhibits outgrowth and promotes branching of
neurites in the presence of inhibitory CNS myelin. FASEB J. 2006; 20:1939–1941. [PubMed:
16849393]
Alele J, Jiang J, Goldsmith JF, Yang X, Maheshwari HG, Black RA, Baumann G, Frank SJ. Blockade
of growth hormone receptor shedding by a metalloprotease inhibitor. Endocrinology. 1998;
139:1927–1935. [PubMed: 9528979]
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 11
Arribas J, Coodly L, Vollmer P, Kishimoto TK, Rose-John S, Massague J. Diverse cell surface protein
ectodomains are shed by a system sensitive to metalloprotease inhibitors. J Biol Chem. 1996;
Author Manuscript
prevents exit from the TGN and proteolytic processing by furin. FEBS Lett. 2001; 505:118–124.
[PubMed: 11557053]
Bazil V, Strominger JL. Metalloprotease and serine protease are involved in cleavage of CD43, CD44,
and CD16 from stimulated human granulocytes. Induction of cleavage of L-selectin via CD16. J
Immunol. 1994; 152:1314–1322. [PubMed: 7507963]
Becker JC, Dummer R, Hartmann AA, Burg G, Schmidt RE. Shedding of ICAM-1 from human
melanoma cell lines induced by IFN-gamma and tumor necrosis factor-alpha. Functional
consequences on cell-mediated cytotoxicity. J Immunol. 1991; 147:4398–4401. [PubMed:
1684377]
Beer S, Oleszewski M, Gutwein P, Geiger C, Altevogt P. Metalloproteinase-mediated release of the
ectodomain of L1 adhesion molecule. J Cell Sci. 1999; 112(Pt 16):2667–2675. [PubMed:
10413675]
Bergmeier W, Rabie T, Strehl A, Piffath CL, Prostredna M, Wagner DD, Nieswandt B. GPVI down-
regulation in murine platelets through metalloproteinase-dependent shedding. Thromb Haemost.
2004; 91:951–958. [PubMed: 15116256]
Author Manuscript
Black RA, Rauch CT, Kozlosky CJ, Peschon JJ, Slack JL, Wolfson MF, Castner BJ, Stocking KL,
Reddy P, Srinivasan S, Nelson N, Boiani N, Schooley KA, Gerhart M, Davis R, Fitzner JN,
Johnson RS, Paxton RJ, March CJ, Cerretti DP. A metalloproteinase disintegrin that releases
tumour-necrosis factor-alpha from cells. Nature. 1997; 385:729–733. [PubMed: 9034190]
Bohlson SS, Silva R, Fonseca MI, Tenner AJ. CD93 is rapidly shed from the surface of human
myeloid cells and the soluble form is detected in human plasma. J Immunol. 2005; 175:1239–
1247. [PubMed: 16002728]
Borrell-Pages M, Rojo F, Albanell J, Baselga J, Arribas J. TACE is required for the activation of the
EGFR by TGF-alpha in tumors. EMBO J. 2003; 22:1114–1124. [PubMed: 12606576]
Borroto A, Ruiz-Paz S, de la Torre TV, Borrell-Pages M, Merlos-Suarez A, Pandiella A, Blobel CP,
Baselga J, Arribas J. Impaired trafficking and activation of tumor necrosis factor-alpha-converting
enzyme in cell mutants defective in protein ectodomain shedding. J Biol Chem. 2003; 278:25933–
25939. [PubMed: 12714588]
Bosenberg MW, Pandiella A, Massague J. The cytoplasmic carboxy-terminal amino acid specifies
Author Manuscript
cleavage of membrane TGF alpha into soluble growth factor. Cell. 1992; 71:1157–1165.
[PubMed: 1473151]
Brachmann R, Lindquist PB, Nagashima M, Kohr W, Lipari T, Napier M, Derynck R. Transmembrane
TGF-alpha precursors activate EGF/TGF-alpha receptors. Cell. 1989; 56:691–700. [PubMed:
2645058]
Brakebusch C, Varfolomeev EE, Batkin M, Wallach D. Structural requirements for inducible shedding
of the p55 tumor necrosis factor receptor. J Biol Chem. 1994; 269:32488–32496. [PubMed:
7798250]
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 12
[PubMed: 8626810]
Croucher PI, Wang F, Hargreaves PG. Interleukin-6 receptor shedding: a possible role for members of
the ADAM family. Biochem Soc Trans. 1999; 27:224–228. [PubMed: 10093738]
Crowe PD, VanArsdale TL, Goodwin RG, Ware CF. Specific induction of 80-kDa tumor necrosis
factor receptor shedding in T lymphocytes involves the cytoplasmic domain and phosphorylation.
J Immunol. 1993; 151:6882–6890. [PubMed: 8258697]
Cui X, Hawari F, Alsaaty S, Lawrence M, Combs CA, Geng W, Rouhani FN, Miskinis D, Levine SJ.
Identification of ARTS-1 as a novel TNFR1-binding protein that promotes TNFR1 ectodomain
shedding. J Clin Invest. 2002; 110:515–526. [PubMed: 12189246]
Cui X, Rouhani FN, Hawari F, Levine SJ. An aminopeptidase, ARTS-1, is required for interleukin-6
receptor shedding. J Biol Chem. 2003a; 278:28677–28685. [PubMed: 12748171]
Cui X, Rouhani FN, Hawari F, Levine SJ. Shedding of the type II IL-1 decoy receptor requires a
multifunctional aminopeptidase, aminopeptidase regulator of TNF receptor type 1 shedding. J
Immunol. 2003b; 171:6814–6819. [PubMed: 14662887]
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 13
regulates angiogenic processes in microvascular endothelial cells. J Biol Chem. 2006; 281:14533–
14536. [PubMed: 16574663]
Fiore E, Fusco C, Romero P, Stamenkovic I. Matrix metalloproteinase 9 (MMP-9/gelatinase B)
proteolytically cleaves ICAM-1 and participates in tumor cell resistance to natural killer cell-
mediated cytotoxicity. Oncogene. 2002; 21:5213–5223. [PubMed: 12149643]
Fitzgerald ML, Wang Z, Park PW, Murphy G, Bernfield M. Shedding of syndecan-1 and -4
ectodomains is regulated by multiple signaling pathways and mediated by a TIMP-3-sensitive
metalloproteinase. J Cell Biol. 2000; 148:811–824. [PubMed: 10684261]
Fourie AM, Coles F, Moreno V, Karlsson L. Catalytic activity of ADAM8, ADAM15, and MDC-L
(ADAM28) on synthetic peptide substrates and in ectodomain cleavage of CD23. J Biol Chem.
2003; 278:30469–30477. [PubMed: 12777399]
Franzke CW, Tasanen K, Schacke H, Zhou Z, Tryggvason K, Mauch C, Zigrino P, Sunnarborg S, Lee
DC, Fahrenholz F, Bruckner-Tuderman L. Transmembrane collagen XVII, an epithelial adhesion
protein, is shed from the cell surface by ADAMs. EMBO J. 2002; 21:5026–5035. [PubMed:
12356719]
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 14
Thomas W, Wells G, Wood LM, Wooley K. Processing of tumour necrosis factor-alpha precursor
by metalloproteinases. Nature. 1994; 370:555–557. [PubMed: 8052310]
Author Manuscript
Gearing AJ, Beckett P, Christodoulou M, Churchill M, Clements JM, Crimmin M, Davidson AH,
Drummond AH, Galloway WA, Gilbert R, Gordon LJ, Leber TM, Mangan M, Miller K, Nayee P,
Owen K, Patel S, Thomas W, Wells G, Wood LM, Wooley K. Matrix metalloproteinases and
processing of pro-TNF-alpha. J Leukoc Biol. 1995; 57:774–777. [PubMed: 7759957]
Gechtman Z, Alonso JL, Raab G, Ingber DE, Klagsbrun M. The shedding of membrane-anchored
heparin-binding epidermal-like growth factor is regulated by the Raf/mitogen-activated protein
kinase cascade and by cell adhesion and spreading. J Biol Chem. 1999; 274:28828–28835.
[PubMed: 10497257]
Gibot S, Kolopp-Sarda MN, Bene MC, Bollaert PE, Lozniewski A, Mory F, Levy B, Faure GC. A
soluble form of the triggering receptor expressed on myeloid cells-1 modulates the inflammatory
response in murine sepsis. J Exp Med. 2004; 200:1419–1426. [PubMed: 15557347]
Gomez-Pina V, Soares-Schanoski A, Rodriguez-Rojas A, Del Fresno C, Garcia F, Vallejo-Cremades
MT, Fernandez-Ruiz I, Arnalich F, Fuentes-Prior P, Lopez-0Collazo E. Metalloproteinases shed
TREM-1 ectodomain from lipopolysaccharide-stimulated human monocytes. J Immunol. 2007;
Author Manuscript
Haro H, Crawford HC, Fingleton B, Shinomiya K, Spengler DM, Matrisian LM. Matrix
metalloproteinase-7-dependent release of tumor necrosis factor-alpha in a model of herniated disc
resorption. J Clin Invest. 2000; 105:143–150. [PubMed: 10642592]
Hattori M, Osterfield M, Flanagan JG. Regulated cleavage of a contact-mediated axon repellent.
Science. 2000; 289:1360–1365. [PubMed: 10958785]
Hayashida K, Kume N, Murase T, Minami M, Nakagawa D, Inada T, Tanaka M, Ueda A, Kominami
G, Kambara H, Kimura T, Kita T. Serum soluble lectin-like oxidized low-density lipoprotein
receptor-1 levels are elevated in acute coronary syndrome: a novel marker for early diagnosis.
Circulation. 2005; 112:812–818. [PubMed: 16061745]
Heissig B, Hattori K, Dias S, Friedrich M, Ferris B, Hackett NR, Crystal RG, Besmer P, Lyden D,
Moore MA, Werb Z, Rafii S. Recruitment of stem and progenitor cells from the bone marrow
niche requires MMP-9 mediated release of kit-ligand. Cell. 2002; 109:625–637. [PubMed:
12062105]
Heiz M, Grunberg J, Schubiger PA, Novak-Hofer I. Hepatocyte growth factor-induced ectodomain
Author Manuscript
shedding of cell adhesion molecule L1: role of the L1 cytoplasmic domain. J Biol Chem. 2004;
279:31149–31156. [PubMed: 15151998]
Herman C, Chernajovsky Y. Mutation of proline 211 reduces shedding of the human p75 TNF
receptor. J Immunol. 1998; 160:2478–2487. [PubMed: 9498793]
Herren B, Levkau B, Raines EW, Ross R. Cleavage of beta-catenin and plakoglobin and shedding of
VE-cadherin during endothelial apoptosis: evidence for a role for caspases and metalloproteinases.
Mol Biol Cell. 1998; 9:1589–1601. [PubMed: 9614196]
Hikita A, Tanaka S. Ectodomain shedding of receptor activator of NF-κB ligand. Adv Exp Med Biol.
2007; 602:15–21. [PubMed: 17966383]
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 15
Hooper NM, Trew AJ, Parkin ET, Turner AJ. The role of proteolysis in Alzheimer’s disease. Adv Exp
Med Biol. 2000; 477:379–390. [PubMed: 10849764]
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 16
Kaup M, Dassler K, Weise C, Fuchs H. Shedding of the transferrin receptor is mediated constitutively
by an integral membrane metalloprotease sensitive to tumor necrosis factor alpha protease
Author Manuscript
secretase and presenilin/gamma-secretase and release signaling fragments. J Biol Chem. 2003;
278:34427–34437. [PubMed: 12826675]
Le Gall SM, Auger R, Dreux C, Mauduit P. Regulated cell surface pro-EGF ectodomain shedding is a
zinc metalloprotease-dependent process. J Biol Chem. 2003; 278:45255–45268. [PubMed:
12947092]
Leduc D, Beaufort N, de Bentzmann S, Rousselle JC, Namane A, Chignard M, Pidard D. The
Pseudomonas aeruginosa LasB metalloproteinase regulates the human urokinase-type
plasminogen activator receptor through domain-specific endoproteolysis. Infect Immun. 2007;
75:3848–3858. [PubMed: 17517866]
Lemjabbar H, Basbaum C. Platelet-activating factor receptor and ADAM10 mediate responses to
Staphylococcus aureus in epithelial cells. Nat Med. 2002; 8:41–46. [PubMed: 11786905]
Letellier M, Nakajima T, Pulido-Cejudo G, Hofstetter H, Delespesse G. Mechanism of formation of
human IgE-binding factors (soluble CD23): III. Evidence for a receptor (Fc epsilon RII)-
associated proteolytic activity. J Exp Med. 1990; 172:693–700. [PubMed: 2143772]
Li N, Wang Y, Forbes K, Vignali KM, Heale BS, Saftig P, Hartmann D, Black RA, Rossi JJ, Blobel
Author Manuscript
CP, Dempsey PJ, Workman CJ, Vignali DA. Metalloproteases regulate T-cell proliferation and
effector function via LAG-3. EMBO J. 2007; 26:494–504. [PubMed: 17245433]
Li Q, Park PW, Wilson CL, Parks WC. Matrilysin shedding of syndecan-1 regulates chemokine
mobilization and transepithelial efflux of neutrophils in acute lung injury. Cell. 2002; 111:635–
646. [PubMed: 12464176]
Lichtenthaler SF. Ectodomain shedding of the amyloid precursor protein: cellular control mechanisms
and novel modifiers. Neurodegener Dis. 2006; 3:262–269. [PubMed: 17047366]
Lichtenthaler SF, Haass C. Amyloid at the cutting edge: activation of alpha-secretase prevents
amyloidogenesis in an Alzheimer disease mouse model. J Clin Invest. 2004; 113:1384–1387.
[PubMed: 15146234]
Lin YZ, Clinton GM. A soluble protein related to the HER-2 proto-oncogene product is released from
human breast carcinoma cells. Oncogene. 1991; 6:639–643. [PubMed: 1674366]
Loechel F, Overgaard MT, Oxvig C, Albrechtsen R, Wewer UM. Regulation of human ADAM 12
protease by the prodomain. Evidence for a functional cysteine switch. J Biol Chem. 1999;
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 17
like protease in shedding of TRANCE, a TNF family member involved in osteoclastogenesis and
dendritic cell survival. J Biol Chem. 1999; 274:13613–13618. [PubMed: 10224132]
Author Manuscript
Madge LA, Sierra-Honigmann MR, Pober JS. Apoptosis-inducing agents cause rapid shedding of
tumor necrosis factor receptor 1 (TNFR1). A nonpharmacological explanation for inhibition of
TNF-mediated activation. J Biol Chem. 1999; 274:13643–13649. [PubMed: 10224136]
Maretzky T, Reiss K, Ludwig A, Buchholz J, Scholz F, Proksch E, de Strooper B, Hartmann D, Saftig
P. ADAM10 mediates E-cadherin shedding and regulates epithelial cell-cell adhesion, migration,
and beta-catenin translocation. Proc Natl Acad Sci USA. 2005; 102:9182–9187. [PubMed:
15958533]
Marron MB, Singh H, Tahir TA, Kavumkal J, Kim HZ, Koh GY, Brindle NP. Regulated proteolytic
processing of Tie1 modulates ligand responsiveness of the receptor-tyrosine kinase Tie2. J Biol
Chem. 2007; 282:30509–30517. [PubMed: 17728252]
Maruyama K, Kametani F, Usami M, Yamao-Harigaya W, Tanaka K. “Secretase,” Alzheimer amyloid
protein precursor secreting enzyme is not sequence-specific. Biochem Biophys Res Commun.
1991; 179:1670–1676. [PubMed: 1930205]
Matala E, Alexander SR, Kishimoto TK, Walcheck B. The cytoplasmic domain of L-selectin
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 18
Mullberg J, Oberthur W, Lottspeich F, Mehl E, Dittrich E, Graeve L, Heinrich PC, Rose-John S. The
soluble human IL-6 receptor. Mutational characterization of the proteolytic cleavage site. J
Author Manuscript
14761956]
Neumann S, Schobel S, Jager S, Trautwein A, Haass C, Pietrzik CU, Lichtenthaler SF. Amyloid
precursor-like protein 1 influences endocytosis and proteolytic processing of the amyloid
precursor protein. J Biol Chem. 2006; 281:7583–7594. [PubMed: 16344553]
Nichols JT, Miyamoto A, Olsen SL, D’Souza B, Yao C, Weinmaster G. DSL ligand endocytosis
physically dissociates Notch1 heterodimers before activating proteolysis can occur. J Cell Biol.
2007; 176:445–458. [PubMed: 17296795]
Noe V, Fingleton B, Jacobs K, Crawford HC, Vermeulen S, Steelant W, Bruyneel E, Matrisian LM,
Mareel M. Release of an invasion promoter E-cadherin fragment by matrilysin and
stromelysin-1. J Cell Sci. 2001; 114:111–118. [PubMed: 11112695]
Ohnishi H, Kobayashi H, Okazawa H, Ohe Y, Tomizawa K, Sato R, Matozaki T. Ectodomain
shedding of SHPS-1 and its role in regulation of cell migration. J Biol Chem. 2004; 279:27878–
27887. [PubMed: 15123722]
Osenkowski P, Toth M, Fridman R. Processing, shedding, and endocytosis of membrane type 1-matrix
metalloproteinase (MT1-MMP). J Cell Physiol. 2004; 200:2–10. [PubMed: 15137052]
Author Manuscript
Pan D, Rubin GM. Kuzbanian controls proteolytic processing of notch and mediates lateral inhibition
during Drosophila and vertebrate neurogenesis. Cell. 1997; 90:271–280. [PubMed: 9244301]
Pandiella A, Bosenberg MW, Huang EJ, Besmer P, Massague J. Cleavage of membrane-anchored
growth factors involves distinct protease activities regulated through common mechanisms. J
Biol Chem. 1992; 267:24028–24033. [PubMed: 1385433]
Park PW, Foster TJ, Nishi E, Duncan SJ, Klagsbrun M, Chen Y. Activation of syndecan-1 ectodomain
shedding by Staphylococcus aureus alpha-toxin and beta-toxin. J Biol Chem. 2004; 279:251–
258. [PubMed: 14573623]
Park PW, Pier GB, Hinkes MT, Bernfield M. Exploitation of syndecan-1 shedding by Pseudomonas
aeruginosa enhances virulence. Nature. 2001; 411:98–102. [PubMed: 11333985]
Park PW, Pier GB, Preston MJ, Goldberger O, Fitzgerald ML, Bernfield M. Syndecan-1 shedding is
enhanced by LasA, a secreted virulence factor of Pseudomonas aeruginosa. J Biol Chem. 2000;
275:3057–3064. [PubMed: 10652286]
Peschon JJ, Slack JL, Reddy P, Stocking KL, Sunnarborg SW, Lee DC, Russell WE, Castner BJ,
Author Manuscript
Johnson RS, Fitzner JN, Boyce RW, Nelson N, Kozlosky CJ, Wolfson MF, Rauch CT, Cerretti
DP, Paxton RJ, March CJ, Black RA. An essential role for ectodomain shedding in mammalian
development. Science. 1998; 282:1281–1284. [PubMed: 9812885]
Phong MC, Gutwein P, Kadel S, Hexel K, Altevogt P, Linderkamp O, Brenner B. Molecular
mechanisms of L-selectin-induced co-localization in rafts and shedding [corrected]. Biochem
Biophys Res Commun. 2003; 300:563–569. [PubMed: 12504120]
Porteu F, Nathan C. Shedding of tumor necrosis factor receptors by activated human neutrophils. J Exp
Med. 1990; 172:599–607. [PubMed: 2165128]
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 19
Powell WC, Fingleton B, Wilson CL, Boothby M, Matrisian LM. The metalloproteinase matrilysin
proteolytically generates active soluble Fas ligand and potentiates epithelial cell apoptosis. Curr
Author Manuscript
Sadhukhan R, Santhamma KR, Reddy P, Peschon JJ, Black RA, Sen I. Unaltered cleavage and
secretion of angiotensin-converting enzyme in tumor necrosis factor-alpha-converting enzyme-
deficient mice. J Biol Chem. 1999; 274:10511–10516. [PubMed: 10187843]
Sahin U, Weskamp G, Kelly K, Zhou HM, Higashiyama S, Peschon J, Hartmann D, Saftig P, Blobel
CP. Distinct roles for ADAM10 and ADAM17 in ectodomain shedding of six EGFR ligands. J
Cell Biol. 2004; 164:769–779. [PubMed: 14993236]
Sanderson MP, Erickson SN, Gough PJ, Garton KJ, Wille PT, Raines EW, Dunbar AJ, Dempsey PJ.
ADAM10 mediates ectodomain shedding of the betacellulin precursor activated by p-
aminophenylmercuric acetate and extracellular calcium influx. J Biol Chem. 2005; 280:1826–
1837. [PubMed: 15507448]
Santhamma KR, Sadhukhan R, Kinter M, Chattopadhyay S, McCue B, Sen I. Role of tyrosine
phosphorylation in the regulation of cleavage secretion of angiotensin-converting enzyme. J Biol
Chem. 2004; 279:40227–40236. [PubMed: 15252021]
Santhamma KR, Sen I. Specific cellular proteins associate with angiotensin-converting enzyme and
Author Manuscript
regulate its intracellular transport and cleavage-secretion. J Biol Chem. 2000; 275:23253–23258.
[PubMed: 10783385]
Schiavo G, van der Goot FG. The bacterial toxin toolkit. Nat Rev Mol Cell Biol. 2001; 2:530–537.
[PubMed: 11433367]
Schlondorff J, Blobel CP. Metalloprotease-disintegrins: modular proteins capable of promoting cell-
cell interactions and triggering signals by protein-ectodomain shedding. J Cell Sci. 1999; 112(Pt
21):3603–3617. [PubMed: 10523497]
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 20
Schlondorff J, Lum L, Blobel CP. Biochemical and pharmacological criteria define two shedding
activities for TRANCE/OPGL that are distinct from the tumor necrosis factor alpha convertase. J
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 21
Tanaka M, Itai T, Adachi M, Nagata S. Downregulation of Fas ligand by shedding. Nat Med. 1998;
4:31–36. [PubMed: 9427603]
Author Manuscript
7919334]
Vaisanen MR, Vaisanen T, Pihlajaniemi T. The shed ectodomain of type XIII collagen affects cell
behaviour in a matrix-dependent manner. Biochem J. 2004; 380:685–693. [PubMed: 15005656]
Veit G, Zimina EP, Franzke CW, Kutsch S, Siebolds U, Gordon MK, Bruckner-Tuderman L, Koch M.
Shedding of collagen XXIII is mediated by furin and depends on the plasma membrane
microenvironment. J Biol Chem. 2007; 282:27424–27435. [PubMed: 17627939]
Velasco-Loyden G, Arribas J, Lopez-Casillas F. The shedding of betaglycan is regulated by
pervanadate and mediated by membrane type matrix metalloprotease-1. J Biol Chem. 2004;
279:7721–7733. [PubMed: 14672946]
Vollmer P, Walev I, Rose-John S, Bhakdi S. Novel pathogenic mechanism of microbial
metalloproteinases: liberation of membrane-anchored molecules in biologically active form
exemplified by studies with the human interleukin-6 receptor. Infect Immun. 1996; 64:3646–
3651. [PubMed: 8751912]
von Bredow DC, Nagle RB, Bowden GT, Cress AE. Cleavage of beta 4 integrin by matrilysin. Exp
Author Manuscript
[PubMed: 16557002]
Weskamp G, Schlondorff J, Lum L, Becherer JD, Kim TW, Saftig P, Hartmann D, Murphy G, Blobel
CP. Evidence for a critical role of the tumor necrosis factor alpha convertase (TACE) in
ectodomain shedding of the p75 neurotrophin receptor (p75NTR). J Biol Chem. 2004; 279:4241–
4249. [PubMed: 14638693]
Wheeler DL, Ness KJ, Oberley TD, Verma AK. Protein kinase Cepsilon is linked to 12-O-
tetradecanoylphorbol-13-acetate-induced tumor necrosis factor-alpha ectodomain shedding and
the development of metastatic squamous cell carcinoma in protein kinase Cepsilon transgenic
mice. Cancer Res. 2003; 63:6547–6555. [PubMed: 14559850]
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 22
Wild-Bode C, Fellerer K, Kugler J, Haass C, Capell A. A basolateral sorting signal directs ADAM10
to adherens junctions and is required for its function in cell migration. J Biol Chem. 2006;
Author Manuscript
Xu J, Park PW, Kheradmand F, Corry DB. Endogenous attenuation of allergic lung inflammation by
syndecan-1. J Immunol. 2005; 174:5758–5765. [PubMed: 15843578]
Yabkowitz R, Meyer S, Black T, Elliott G, Merewether LA, Yamane HK. Inflammatory cytokines and
vascular endothelial growth factor stimulate the release of soluble tie receptor from human
endothelial cells via metalloprotease activation. Blood. 1999; 93:1969–1979. [PubMed:
10068670]
Yu WH, Woessner JF Jr. McNeish JD, Stamenkovic I. CD44 anchors the assembly of matrilysin/
MMP-7 with heparin-binding epidermal growth factor precursor and ErbB4 and regulates female
reproductive organ remodeling. Genes Dev. 2002; 16:307–323. [PubMed: 11825873]
Zhang Z, Oliver P, Lancaster JJ, Schwarzenberger PO, Joshi MS, Cork J, Kolls JK. Reactive oxygen
species mediate tumor necrosis factor alpha-converting, enzyme-dependent ectodomain shedding
induced by phorbol myristate acetate. FASEB J. 2001; 15:303–305. [PubMed: 11156944]
Zhao J, Chen H, Peschon JJ, Shi W, Zhang Y, Frank SJ, Warburton D. Pulmonary hypoplasia in mice
lacking tumor necrosis factor-alpha converting enzyme indicates an indispensable role for cell
surface protein shedding during embryonic lung branching morphogenesis. Dev Biol. 2001a;
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 23
Author Manuscript
Author Manuscript
Author Manuscript
Fig. 1.
Regulatory mechanisms of ectodomain shedding. Mechanisms as diverse as protein–protein
interactions, phosphorylation, intracellular trafficking, polarized secretion, and activation of
sheddases contribute to the regulation of ectodomain shedding at the cell surface. Several
examples are shown. (1) Intracellular protein–protein interaction: Calmodulin constitutively
bound to the cytoplasmic tail of substrates (e.g., L-selectin, ACE) inhibits ectodomain
shedding, and the dissociation of calmodulin induced by calmodulin kinase enhances
shedding. In contrast, PMA stimulation induces the association of moesin, potentiating the
shedding of substrates (e.g., L-selectin). (2) Extracellular protein–protein interaction:
Binding of ARTS-1 to cytokine receptors, such as TNFRI and IL-6R, activates shedding
possibly by inducing a conformational change in the substrate or by displacing an inhibitory
factor from the substrate. (3) Intracellular trafficking of substrate: BiP binds to substrates
Author Manuscript
(e.g., ACE) and retains the substrate in the ER, preventing its encounter with the sheddase at
the cell surface. (4) Phosphorylation of sheddase: PTKs or PKCs may activate the sheddase
through Tyr or Ser/Thr phosphorylation of the cytoplasmic tail of the membrane-associated
sheddase. (5) Activation of sheddase: Sheddases belonging to the ADAM or MMP family
are activated by removal of the prodomain by furin and furin-like PCs in the trans Golgi
compartment and also at the cell surface. (6) Intracellular trafficking of sheddase: TACE/
ADAM17 is trafficked to the cell surface in a phosphorylation-dependent manner. For
example, gastrin-releasing peptide activates a Src-PI3K-PDK pathway that induces Ser/Thr
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 24
phosphorylation and promotes ADAM translocation to the cell surface. (7) Mobilization to
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.
HAYASHIDA et al. Page 25
TABLE 1
Partial list of proteins released from the cell surface by ectodomain shedding
Author Manuscript
Cell adhesion molecules Growth factors and receptors Immunomodulators and receptors
Collagen XIII (Vaisanen et al., 2004) Amphiregulin (Sahin et al., 2004) Betaglycan (Velasco-Loyden et al., 2004)
Collagen XVII (Franzke et al., 2002) Betacellulin (Sahin et al., 2004) CD23 (Letellier et al., 1990)
Collagen XXIII (Veit et al., 2007) CHL1 (Naus et al., 2004) CD30 (Hansen et al., 2004)
E-Cadherin (McGuire et al., 2003) c-Met (Nath et al., 2001) CD40 (Contin et al., 2003)
N-Cadherin (Reiss et al., 2005) EGF (Sahin et al., 2004) CD93 (Bohlson et al., 2005)
VE-Cadherin (Herren et al., 1998) EGFRs (Lin and Clinton, 1991) CD223 (Li et al., 2007)
CD44 (Bazil and Strominger, 1994) Ephrins (Hattori et al., 2000) CSF-1 (Tuck et al., 1994)
Glycoprotein V (Rabie et al., 2005) Epiregulin (Sahin et al., 2004) CXCL16 (Matloubian et al., 2000)
Glycoprotein VI (Bergmeier et al., 2004) FGFR1 (Hanneken et al., 1994) Fas (Cheng et al., 1994)
ICAM1 (Becker et al., 1991) GHR (Alele et al., 1998) FasL (Tanaka et al., 1998)
Author Manuscript
L1 (Beer et al., 1999) HB-EGF (Suzuki et al., 1997) Fractalkine (Garton et al., 2001)
NCAM (Hubschmann et al., 2005) HGF (Mizuno et al., 1994) GM-CSFR (Prevost et al., 2002)
Nectin-1α (Tanaka et al., 2002) KL-1 (Pandiella et al., 1992) IL-1RII (Cui et al., 2003b)
Nectin-4 (Fabre-Lafay et al., 2005) Neuregulin (Sahin et al., 2004) IL-2R (Junghans and Waldmann, 1996)
Neuroglycan C (Shuo et al., 2007) NgR (Ahmed et al., 2006) IL-6R (Croucher et al., 1999)
NG2 (Asher et al., 2005) NTR(p75) (Weskamp et al., 2004) IL-15R (Mortier et al., 2004)
PECAM-1 (Ilan et al., 2001) Osteoactivin (Furochi et al., 2007) LAR (Streuli et al., 1992)
E-selectin (Wyble et al., 1997) TGFα (Wong et al., 1989) LOX-1 (Hayashida et al., 2005)
L-selectin (Migaki et al., 1995) TIE1 (Marron et al., 2007) RANKL (Hikita and Tanaka, 2007)
P-selectin (Semenov et al., 1999) TSHR (Couet et al., 1996) TNFα (Black et al., 1997)
SHPS-1 (Ohnishi et al., 2004) TNFRI, TNFRII (Porteu and Nathan, 1990)
Syndecan-1 (Subramanian et al., 1997) TRANCE (Lum et al., 1999)
Author Manuscript
Anat Rec (Hoboken). Author manuscript; available in PMC 2015 October 27.