LETTER TO THE EDITOR
Apo-RBP, Holo-RBP, and Insulin Resistance
Ming Zhong, Riki Kawaguchi, Miki Kassai, Hui Sun
Department of Physiology, Jules Stein Eye Institute, and Howard Hughes Medical Institute, David Geffen School of Medicine, University of California, Los Angeles,
California, USA
B erry et al. claimed that STRA6 binds only holo-retinol/retinol
binding protein complex (holo-RBP), not apo-RBP, and that
holo-RBP’s binding to STRA6 causes insulin resistance (1). These
claims are exactly the opposite of the conclusions from two studies
in Molecular and Cellular Biology by independent groups who
showed that RBP’s effect on insulin resistance does not depend
on STRA6 and that apo-RBP triggers insulin resistance (2, 3).
Contrary to the claim by Berry et al., obesity is associated with
significantly higher apo-RBP, but normal holo-RBP (4), and
that the RBP-to-retinol ratio is elevated in patients with type 2
diabetes (5).
The claims by Berry et al. contradict not only findings from
other labs on RBP but also known properties of STRA6. Contrary
to the claim by Berry et al., apo-RBP does bind to STRA6. First,
both Isken and colleagues (6) and we (7) showed that apo-RBP
can promote retinol efflux through STRA6. Second, it has been
shown that both apo-RBP and holo-RBP bind to STRA6 (7).
Third, STRA6’s abilities to bind holo-RBP and apo-RBP make it
possible for STRA6 to catalyze retinol release from holo-RBP and
to catalyze retinol loading into apo-RBP (8). Fourth, in contrast to
STRA6’s role in insulin resistance, STRA6 expression is highly
decreased in adipose tissues in wild-type mice and ob/ob mice fed
a high-fat diet (9). Fifth, Berry et al. initially linked STRA6 to
insulin resistance by purifying human STRA6 using a commercial
antibody that does not recognize human STRA6, as shown by two
FIG 1 [3H]retinol uptake assay from [3H]retinol/RBP. Purified [3H]retinol/
RBP was incubated with cells for 1 h. Like wild-type STRA6 (STRA6-WT), the
L255A mutant of STRA6 (STRA6-L255A) couples to CRBP-I or LRAT for
vitamin A uptake from holo-RBP. The negative control represents RBP recep-
tor-independent and nonspecific binding of [3H]retinol/RBP to plastic dish or
cells.
June 2014 Volume 34 Number 11 Molecular and Cellular Biology
independent studies (7, 10). Consistently, the so-called STRA6
bands never increased in intensity after STRA6 transfection (11).
Surprisingly, Berry et al. did not do any experiment but solely
relied on online computer software, SOSUI, to determine which
STRA6 residues are intracellular or extracellular and to conclude
that STRA6 has 11 transmembrane domains. A study 6 years ago
experimentally determined that the 11-transmembrane domain
topology is incorrect (12). Computer prediction is both unreliable
and uncertain. For example, SOSUI predicts mouse STRA6 as a
10-transmembrane domain protein and zebrafish STRA6 as a
6-transmembrane domain protein.
Contrary to the claim by Berry et al., STRA6 does not strictly
depend on cellular retinol binding protein I (CRBP-I) for its ac-
tivity. We and other investigators showed that either lecithin ret-
inol acyltransferase (LRAT) (6, 8, 13–15) or CRBP-I (8) can stim-
ulate STRA6’s vitamin A uptake activity from holo-RBP. STRA6
also has many LRAT- or CRBP-I-independent activities, and
many of these activities are suppressed, not enhanced, by the pres-
ence of either CRBP-I or LRAT (7, 8). For example, STRA6 cata-
lyzes retinol release from holo-RBP independently of LRAT and
CRBP-I. STRA6 also catalyzes retinol loading into apo-RBP inde-
pendently of LRAT and CRBP-I. Not only does STRA6 not de-
pend on LRAT or CRBP-I for this activity, LRAT or CRBP-I actu-
ally inhibits STRA6’s activity. STRA6 also catalyzes retinol
exchange between RBP molecules independently of LRAT or
CRBP-I (7). Like LRAT or CRBP-I, other proteins or even small
molecules like ␤-ionone can enhance STRA6’s vitamin A uptake
activity (8). We have tested the STRA6 residue claimed by Berry et
al. to be essential for STRA6/CRBP-I interaction and found that its
mutation (L255A) does not affect STRA6’s coupling to either
CRBP-I or LRAT for vitamin A uptake from holo-RBP (Fig. 1).
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Address correspondence to Hui Sun, hsun@mednet.ucla.edu.
For the author reply, see doi:10.1128/MCB.01699-12.
Copyright © 2014, American Society for Microbiology. All Rights Reserved.
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Molecular and Cellular Biology