 Protein product of ABHD5(Alpha Beta Hydrolase Domain 5): 1-

acylglycerol-3-phosphate O-acyltransferase ABHD5

 Localisation in cell: cytoplasm and surface of lipid droplets (LDs) (of

adiopocytes?), localized to LGs in keratinocytes

 Alpha/beta hydrolase superfamily: characterized by an α/β

hydrolase fold domain on (amino acid residues 102–345). Other
members include glutactin, vitellogenin, neurotactin, and
thyroglobulin.

 The esterase/lipase/thioesterase subfamily: determined by order

of catalytic triad motifs. Suggests participation in lipid metabolism and
thus association with disease phenotypes .

https://pure.rug.nl/ws/portalfiles/portal/3321987/1992ProteinEngOllis.pdf

 Functional Structure/domain of ABHD5 protein:

 Catalytic triad(amino acid residues 76–178): the active site which is
composed of a nucleophile, an acid, and histidine(which act as the
base as it is positively charged) that are distant from each other in
sequence but close in three-dimensional structure, corresponding to
the subfamily
 Nuc: Usually the nucleophile should be serine, but it is replaced by
asparagine in ABHD5 protein. Defined by LLGHNLGG motif, amino
acid residues 149–156 (GXSXG motif?)

 Acid: could be either the aspartate residue at 301, inside the motif
GARSCIDG (amino acid residues 295–302), or the glutamic acid
residue at 260, which we found mutated in one family, inside the
motif PSGETA (amino acid residues 257–262).
 Base: histidine at residue 327, inside the motif GsmXHsmX (amino
acid residues 324–328), where “sm” denotes a residue with a small
side chain
 Mechanism of catalytic triad: The side-chain of the nucleophilic
residue performs covalent catalysis on the substrate. The lone pair of
electrons present on the oxygen or sulfur attacks the
electropositive carbonyl carbon.[3] The acid residue
(commonly glutamate or aspartate) aligns and polarises the base
(usually histidine) which activates the nucleophile (often serine or
cysteine, occasionally threonine). The triad reduces the pKa of the
nucleophilic residue which then attacks the substrate.

https://en.wikipedia.org
/wiki/Catalytic_triad

 Interactome: abhd5 protein was shown to interact with

perilipin(which restricts lipolysis) and adipocyte triglyceride lipase
(ATGL) on the LD surface
1. perilipin:
/switching between lipid storage and utilization
/perilipin blocks the access of HSL to LDs in quiescent
adipocytes /and thus restricts hydrolytic attack
/multiphosphorylated perilipin loses its barrier function but
facilitates the access of HSL to LDs, thereby promoting lipolysis
2. CGI-58: A Binding Partner of Perilipin:
/induced during adipocyte differentiation
/In quiescent adipocytes, CGI-58 is colocalized with perilipin on
the surface of LDs. Upon lipolytic stimulation with catecholamines,
CGI-58 is rapidly released from LDs into the cytosol along with
 small particulate structures. This is probably because CGI-58
does not bind to phosphorylated perilipin
/siRNA-mediated knockdown of CGI-58 caused abnormal
accumulation of LDs
/These results indicate that CGI-58 is a lipolytic factor that
facilitates lipolysis,

3. Ichthyosis : results from the malformation of a permeability barrier

in the skin. Truncation of CGI-58 protein results in abnormal
lamellar granule (LG) formation (involved in lipid transport and
secretion in keratinocytes and have an important role in forming
the epidermal lipid barrier). CGI-58 on LGs in keratinocytes and its
expression is upregulated during keratinocyte differentiation and
development of the skin in humans.

/ CGI-58 knockdown reduces the expression of keratinocyte

differentiation markers

/CGI-58 plays crucial roles in epidermal keratinocyte differentiation

and LG lipid metabolism, thereby contributing to the formation of
the skin’s lipid barrier.

/A lesion of phospholipid synthesis from TG was observed in CDS

fibroblasts, and CGI-58 was suggested to be involved in the
acylation of lysophosphatidic acid.( there is a consensus
sequence for acyltransferase activity of HXXXXD near the
carboxyl terminus of all vertebrate CGI-58 sequences.)

/Overexpression of CGI-58 in Saccharomyces cerevisiae led to an

increase in the formation of phosphatidic acid, resulting in an
overall increase in total phospholipids, but the TG level was
significantly reduced.

4. ATGL: A TG Lipase Responsible for Adipocyte Lipolysis

/localized in the cytosol and on the surface of LDs
/ATGL is predominantly expressed in adipose tissue and
catalyzes the hydrolysis of TG to DG and FA.
/Function slightly overlapps with HSL
/ATGL requires CGI-58 for efficient enzyme function. Although
CGI-58 itself does not have lipase activity, it interacts with ATGL
and activates the enzyme activity of the latter in vitro. Thus, CGI-
58 functions as a protein cofactor of ATGL, and mutant forms of
CGI-58 associated with CDS lose the capacity to activate ATGL
  Summary of interaction in lipolysis

Functional Interplay of Proteins on Adipocyte LDs:

The current view of the mechanism of regulation of lipolysis

can be summarized as follows. In quiescent adipocytes,
perilipin binds to CGI-58 on the surface of LDs, hence
preventing CGI-58 from interacting with ATGL. Upon
lipolytic stimulation by catecholamines, PKA phosphorylates
perilipin, and phosphorylated perilipin releases CGI-58. CGI-
58 is now free to interact with ATGL, and the resulting
ATGL/CGI-58 complex efficiently degrades TG to DG and
FA. DG is then hydrolyzed to monoacylglycerol (MG) and
FA by HSL, which is translocated to the LD surface upon
lipolytic stimulation. MG is further decomposed to glycerol
and FA by monoglyceride lipase, and the reaction products
finally enter the circulation

Recent reports revealed that CGI-58 facilitates the use of

lipid intermediates derived from the lipolysis of stored TG for
the assembly of lipoproteins

https://en.wikipedia.org/wiki/File:Catalytic_triad_of_TEV_protease.png
----------------------------------------------------------------------------------------------------

α/β hydrolase fold  (amino acid residues 102–345)

lipases and esterases—implicated in lipid metabolism

(Ollis et al. 
Ollis et al., 1992
; Cygler et al. 
Cygler et al., 1993
; Schrag and Cygler 
Schrag and Cygler, 1997
)

CGI-58 is widely expressed in various tissues, including skin, lymphocytes,

liver, skeletal muscle, and brain (SAGEmap). This broad pattern of
expression is consistent with the CDS phenotype.

The CGI-58 open reading frame encodes a 349-amino-acid protein of ∼39

kD.

cytoplasmic localization for the protein

The strongest homology found was for the sequence motif (amino acid
residues 76–178) bearing one of the three canonical residues that define
the catalytic triad formed by the nucleophile/acid/histidine residues, which
are present in this order in members of the esterase/lipase/thioesterase
subfamily.

An LLGHNLGG motif (the nucleophile component of which is included in

amino acid residues 149–156), in which asparagine replaces serine, is
close to a lipase consensus sequence (LLGHSLGG), in which serine
defines the nucleophilic elbow (Schrag and Cygler 
Schrag and Cygler, 1997
);

serine and asparagine are both polar amino acids

A diagnosis of CDS is easily confirmed by a simple blood smear, in which

the characteristic lipid droplets are observed in the cytoplasm of
granulocytes. These droplets are also seen in leukocytes of heterozygous
carriers of CDS (Wollenberg et al. 
Wollenberg et al., 2000
).

Interactome

Another interesting protein annotated as cgi-58 or abhd5 (α/β hydrolase domain-

containing protein 5) was shown to interact with perilipin(restricts lipolysis) and
adipocyte triglyceride lipase (ATGL) on the LD surface

https://www.jstage.jst.go.jp/article/bpb/33/3/33_3_342/_pdf/-char/en

perilipin:
switching between lipid storage and utilization

perilipin blocks the access of HSL to LDs in quiescent adipocytes and thus restricts
hydrolytic attack

multiphosphorylated perilipin loses its barrier function but facilitates the access of HSL to
LDs, thereby promoting lipolysis

CGI-58: A Binding Partner of Perilipin:

induced during adipocyte differentiation

In quiescent adipocytes, CGI-58 is colocalized with perilipin on the surface of LDs. Upon
lipolytic stimulation with catecholamines, CGI-58 is rapidly released from LDs into the
cytosol along with small particulate structures. This is probably because CGI-58 does not
bind to phosphorylated perilipin

siRNA-mediated knockdown of CGI-58 caused abnormal accumulation of LDs

These results indicate that CGI-58 is a lipolytic factor that facilitates lipolysis,

Ichthyosis : results from the malformation of a permeability barrier in the skin. Truncation of
CGI-58 protein results in abnormal lamellar granule (LG) formation (involved in lipid
transport and secretion in keratinocytes and have an important role in forming the epidermal
lipid barrier). CGI-58 on LGs in keratinocytes and its expression is upregulated during
keratinocyte differentiation and development of the skin in humans.

CGI-58 knockdown reduces the expression of keratinocyte differentiation markers

CGI-58 plays crucial roles in epidermal keratinocyte differentiation and LG lipid metabolism,
thereby contributing to the formation of the skin’s lipid barrier. 28) Brown J. M., Chung S.,
Das A., Shelness G. S., Rudel L. L., Yu L., J. Lipid Res., 48, 2295—2305 (2007). 29)
Caviglia J. M., Sparks J. D., Toraskar N., Brinker A. M., Yin T. C., Dixon J.
A lesion of phospholipid synthesis from TG was observed in CDS fibroblasts, and CGI-58
was suggested to be involved in the acylation of lysophosphatidic acid.( there is a consensus
sequence for acyltransferase activity of HXXXXD near the carboxyl terminus of all
vertebrate CGI-58 sequences.) 25) Igal R. A., Coleman R. A., J. Biol. Chem., 271, 16644—
16651 (1996). 26) Ghosh A. K., Ramakrishnan G., Chandramohan C., Rajasekharan R., J.
Biol. Chem., 283, 24525—24533 (2008). 27) Montero-Moran G. C. J., McMahon D.,
Rothenberg A., Subramanian V., Xu Z., Lara-Gonzalez S., Storch J., Carman G. M.,
Brasaemle D. L., J. Lipid Res., in press (2009).

Overexpression of CGI-58 in Saccharomyces cerevisiae led to an increase in the formation of

phosphatidic acid, resulting in an overall increase in total phospholipids, but the TG level was
significantly reduced. 26) Ghosh A. K., Ramakrishnan G., Chandramohan C., Rajasekharan
R., J. Biol. Chem., 283, 24525—24533 (2008)

ATGL: A TG Lipase Responsible for Adipocyte Lipolysis

localized in the cytosol and on the surface of LDs

ATGL is predominantly expressed in adipose tissue and catalyzes the hydrolysis of TG to DG

and FA.

Function slightly overlapps with HSL

ATGL requires CGI-58 for efficient enzyme function. Although CGI-58 itself does not have
lipase activity, it interacts with ATGL and activates the enzyme activity of the latter in vitro.
Thus, CGI-58 functions as a protein cofactor of ATGL, and mutant forms of CGI-58
associated with CDS lose the capacity to activate ATGL

Functional Interplay of Proteins on Adipocyte LDs:

The current view of the mechanism of regulation of lipolysis can be summarized as follows.
In quiescent adipocytes, perilipin binds to CGI-58 on the surface of LDs, hence preventing
CGI-58 from interacting with ATGL. Upon lipolytic stimulation by catecholamines, PKA
phosphorylates perilipin, and phosphorylated perilipin releases CGI-58. CGI-58 is now free
to interact with ATGL, and the resulting ATGL/CGI-58 complex efficiently degrades TG to
DG and FA. DG is then hydrolyzed to monoacylglycerol (MG) and FA by HSL, which is
translocated to the LD surface upon lipolytic stimulation. MG is further decomposed to
glycerol and FA by monoglyceride lipase, and the reaction products finally enter the
circulation
Recent reports revealed that CGI-58 facilitates the use of lipid intermediates derived from the
lipolysis of stored TG for the assembly of lipoproteins