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ApoB SURFs a Ride from the ER to the Golgi
Henry N. Ginsberg1,*
1Department of Medicine, Vagelos College of Physicians and Surgeons of Columbia University, New York, NY, USA
*Correspondence: hng1@cumc.columbia.edu
https://doi.org/10.1016/j.cmet.2021.01.007
Chylomicrons and very-low-density lipoproteins (VLDLs) are large, complex cargos that may require specific
chaperones for efficient transport from the ER to Golgi. In this issue of Cell Metabolism, Wang et al. (2020)
identify SURF4, in coordination with SAR1B, as an essential player in COPII transport of VLDLs from ER to
Golgi, suggesting that SURF4 may be a target for approaches aimed at reducing secretion of triglyceride-
rich, atherogenic lipoproteins from the liver.
Very-low-density lipoproteins (VLDLs) are
spherical aggregates comprised of a core
of thousands of triglyceride (TG) and
cholesterol ester molecules with a mono-
layer cover of amphipathic phospholipids,
and a small quantity of free cholesterol.
VLDLs are also comprised of several pro-
teins, including apolipoprotein B100
(hereafter referred to as apoB), which is
required for the proper assembly and
secretion of VLDLs. ApoB is comprised
of 4,536 amino acids, including a globular
amino-terminal domain, two very large
highly lipophilic domains separated by a
short amphipathic region, and an amphi-
pathic carboxy-terminal domain. Studies
of the complicated intracellular itinerary
of this very complex polypeptide emerged
in the second half of the 1980s and, for the
next decade, focused on co-translational
lipidation of apoB during its translocation
across the ER membrane and co-translo-
cational proteasomal degradation of
apoB when lipidation was inadequate.
Other types of ER-associated degrada-
tion (ERAD) were also identified during
this time (Fisher and Ginsberg, 2002). In
the first decades of this century, charac-
terization of additional components of
apoB’s itinerary, including post-ER prese-
cretory proteolysis (PERPP), stepwise lip-
idation converting nascent lipid-poor
VLDL that initially enters the ER lumen
into the mature lipid-rich VLDL that is
secreted, and the sites of that lipidation,
have expanded our understanding of the
early trafficking of VLDL (Olofsson and
Borén, 2012).
A less well-defined component of
VLDL’s cellular itinerary, despite more
than a decade of investigation, is its trans-
port from the ER to the Golgi. Although the
COPII vesicular transport system has
been extensively characterized, there
has been uncertainty whether a cargo as
large as VLDL (diameters range from 30
to 100 nm) can ‘‘fit’’ into canonical COPII
vesicles (diameter: 55–80 nm), or require
additional proteins that would facilitate
generation of larger COPII vesicles (Brod-
sky et al., 2004). In a new study in this
issue of Cell Metabolism, Wang et al.
(2020) used an impressive array of ap-
proaches to identify a role for SURF4 as
a unique component of COPII vesicles,
both for carrying VLDL out of the ER and
targeting the vesicle for retrieval from the
Golgi back to the ER. They began their
studies examining the effect of liver-spe-
cific knockout of the cytosolic protein
SAR1B, a well-characterized member of
the Sar1-ADP-ribosylation factor family
of small GTPases that play early, impor-
tant roles in the formation of COPII vesi-
cles. SAR1B is critical for the formation
of very large COPII vesicles in the small in-
testine that transport chylomicrons (diam-
eters ranging from 70 to 600 nm) from the
ER to the Golgi in enterocytes. Loss-of-
function mutations of SAR1B lead to
chylomicron retention disease, also
known as Anderson’s disease, which is
characterized by the absence of circu-
lating chylomicrons, severe fat malab-
sorption, and failure to thrive (Peretti
et al., 2009).
The authors established an SAR1B
liver-specific knockout mouse and
demonstrated significant defects in TG
and apoB secretion, hepatic steatosis,
and marked reductions in plasma levels
of cholesterol, TG, apoB, and apoA-I.
The effects of SAR1B loss of function ap-
peared to be specific for VLDL, as plasma
levels of albumin and alpha1-antitrypsin
were unaffected. The SAR1B knockout
phenotype confirmed reductions in
plasma lipids and hepatic lipoproteins
Cell Metabolism 33, February 2, 2021 ª 2021 Elsevier Inc. 231
present in patients with Anderson’s dis-
ease, and was consistent with stable
isotope kinetics studies in such patients
that demonstrated a 60% reduction in
VLDL apoB secretion and a 30% reduc-
tion in HDL apoA-I secretion (Ouguerram
et al., 2012).
Wang and colleagues next identified
SURF4 as an SAR1B partner using a
cell-based proximity-proteomic assay.
SURF4 has 260 amino acids comprising
6 predicted transmembrane domains,
including one large ER luminal loop,
and a cytosolic sorting motif for ER
retrieval. It is the human homolog of
SFT-4 in C. elegans, which is necessary
for the ER export of yolk proteins, such
as VIT-2, and cargo receptors of the
Erv29p family that bind soluble cargo
and COPII components in yeast.
Knockout of SFT-4 resulted in reduced
ER exit of VIT-2 in C. elegans, and
knockdown of SURF4 in HepG2 cells
led to reduced numbers of ER exit sites
and decreased secretion of apoB (Sae-
gusa et al., 2018). Wang et al. generated
a liver-specific knockout of SURF4,
which resulted in near total knockdown
of plasma TG and cholesterol in all lipo-
protein fractions, concomitant with inhi-
bition of TG secretion and significant he-
patic steatosis. Of note, these mice
showed no evidence of nonalcoholic
steatohepatitis (NASH), including no
fibrosis, at 13 months of age. Transmis-
sion EM studies from livers of the
knockout mice demonstrated accumula-
tion of lipoproteins in the ER and their
absence in the Golgi. The authors pre-
sented immuno-EM using an anti-apoB
antibody, but these difficult studies
were not convincing. Thus, it is uncertain
if the lipoproteins seen with TEM were
apoB-lipoproteins or just ER-lumenal
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cLD cLD
TM6SF2 SAR1A
DGAT1 DGAT2 ERLIN1/2
COPII
PLCE TG
Nascent
TG MTP TG MTP erLD SAR1B
apoB
PL
CE
VLDL VLDL
ER lumen Lipid-poor
VLDL
Lipid-rich
COPII
VLDL VLDL
ER membrane
KLHL12
TANGO1
COPII
Outer nuclear
membrane SAR1A

TorsinA
LAP1

Inner nuclear
membrane Golgi
Nucleus
Figure 1. Proposed Cellular Itinerary of VLDL apoB as It Is Lipidated and Transported from the ER to the Golgi
Despite a number of uncertainties, a generally accepted model of the assembly of VLDL and its transit through the ER to the Golgi includes the following steps:
The initial assembly of VLDL occurs as nascent apoB is lipidated by MTP-mediated transfer of ER-membrane phospholipids, triglycerides, and cholesterol esters
to apoB as it undergoes co-translational translocation into the ER lumen. This relatively lipid-poor VLDL gradually accumulates additional lipids while it remains in
close association with the inner leaflet of the ER membrane, and/or by a physical interaction with lipid droplets within the ER lumen (erLD). MTP appears to be
required for both of these pathways. ERLIN1/2, TM6SF2, and the AAA-ATPase, TorsinA, are all required for VLDL to achieve maturity and be secreted as a
triglyceride-rich apoB lipoprotein. Because VLDLs that are maximally enriched with TG have larger diameters than typical COPII vesicles, modification of these
vesicles is required. Previous studies have identified TANGO1, KHLH12, and SAR1B as proteins that enable COPII vesicle to reach a size that can accommodate
large apoB lipoproteins. SURF4 now joins this select group as a protein that, in collaboration with SAR1B, is necessary for the transfer of large VLDLs from the ER
to the Golgi.

lipid droplets. The authors also pre-
sented immuno-histologic evidence that
apoA-I and apoB accumulate co-inci-
dentally in the ER. Of note, despite these
marked alterations in lipoprotein trans-
port with ER accumulation of lipids,
probably as VLDL, there was no evi-
dence of ER stress or ER autophagy. In
a final series of experiments, Wang and
colleagues demonstrated that (1) bidirec-
tional flux of SURF4 from ER to Golgi and
back to ER was required for the protein’s
full impact on VLDL secretion; (2) SAR1B
and SURF4 acted sequentially to trans-
port VLDL from ER to Golgi; (3) the two
proteins were synergistic, at least in
terms of the effects of heterozygous
loss of function and the hypolipidemic
phenotype; and (4) SURF4 loss of func-
tion protected mice with reduced LDL re-
ceptors from developing atherosclerosis.
What do these very interesting studies
mean for our understanding of the COPII
vesicular transport system from ER to
Golgi? While they clearly add another
player to the team of proteins involved,
they also leave several important ques-
tions unanswered. First, SAR1B is an
early and critical player in COPII vesicle
formation, but the SAR1B loss-of-func-
tion phenotype in humans seems to result
in relatively isolated defects in transport of
large TG-rich lipoproteins. Is this related
to the interaction of SURF4 with SAR1B,
but not SAR1A, which is thought to be
more involved in the generation of smaller
COPIII vesicles that carry typical protein
cargos (Melville et al., 2020)? Second, if
SURF4 activity is key to formation of
larger COPII vesicles, does it interact
with KHLH12 or TANGO1 proteins, also
identified as regulating the size of COPII
vesicles (McCaughey and Stephens,
2018)? Third, is SURF4 the only protein
that can target Golgi COPII vesicle back
to the ER? Finally, where do proteins
such as the ER transmembrane proteins
TM6SF2 and ERLIN (Li et al., 2020), or
TorsinA (Shin et al., 2019), a nuclear mem-
brane-associated AAA ATPase, which
appear to be critical for the assembly
and secretion of mature VLDL, sit along
the apoB secretory highway, and do
they interact with the SAR1B-SURF4
complex (Figure 1)?
The findings of Wang and colleagues
also suggest that targeting hepatic
SURF4 in humans might reduce plasma
lipid levels, thereby reducing cardiovas-
cular disease. Unfortunately, their overall
results suggest that knockdown of
SURF4 will result in hepatic steatosis
that, despite some contrasting data in
this paper, may increase the risk for
developing NASH and cirrhosis, as has
been observed with loss-of-function mu-
tations in apoB or microsomal triglyceride
transfer protein that reduce VLDL assem-
bly and secretion.

232 Cell Metabolism 33, February 2, 2021


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ACKNOWLEDGMENTS
This work was funded by NIH:NHLBI R35
HL135833 to H.N.G. Dr. J.-Y. Shin kindly assisted
with the development of the figure.
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