Abstract
Dietary lipid absorption is dependent on chylomicron production whose rate-limiting step across the intestinal absorptive cell is the exit of chylomicrons from the endoplasmic reticulum (ER) in its ER-to-Golgi transport vesicle, the prechylomicron transport vesicle (PCTV). This study addresses the composition of the budding complex for PCTV. Immunoprecipitation (IP) studies from rat intestinal ER solubilized in Triton X-100 suggested that vesicle-associated membrane protein 7 (VAMP7), apolipoprotein B48 (apoB48), liver fatty acid-binding protein (L-FABP), CD36, and the COPII proteins were associated on incubation of the ER with cytosol and ATP. This association was confirmed by chromatography of the solubilized ER over Sephacryl S400-HR in which these constituents cochromatographed with an apparent kDa of 630. No multiprotein complex was detected when the ER was chromatographed in the absence of PCTV budding activity (resting ER or PKCzeta depletion of ER and cytosol). Treatment of the ER with anti-apoB48 or anti-VAMP7 antibodies or using gene disrupted L-FABP or CD36 mice all significantly inhibited PCTV generation. A smaller complex (no COPII proteins) was formed when only rL-FABP was used to bud PCTV. The data support the conclusion that the PCTV budding complex in intestinal ER is composed of VAMP7, apoB48, CD36, and L-FABP, plus the COPII proteins.
Highlights
Dietary lipid absorption is dependent on chylomicron production whose rate-limiting step across the intestinal absorptive cell is the exit of chylomicrons from the endoplasmic reticulum (ER) in its ER-to-Golgi transport vesicle, the prechylomicron transport vesicle (PCTV)
We showed that the small Mr protein, liver fatty acid-binding protein (L-FABP) (14.1 kDa), can generate cargo-carrying PCTV, the vesicle produced is incomplete in that it cannot fuse with the Golgi (13)
The experiments were based on using three known components of PCTV budding; namely, the unique cargo protein, apolipoprotein B48, which is present in all chylomicrons (18); vesicleassociated membrane protein 7 (VAMP7); the vesicle-associated soluble N-ethylmaleimide-sensitive factor attachment protein receptor (v-SNARE) for PCTV (19); and L-FABP, which as we showed, is able to initiate PCTV budding (13)
Summary
Dietary lipid absorption is dependent on chylomicron production whose rate-limiting step across the intestinal absorptive cell is the exit of chylomicrons from the endoplasmic reticulum (ER) in its ER-to-Golgi transport vesicle, the prechylomicron transport vesicle (PCTV). Immunoprecipitation (IP) studies from rat intestinal ER solubilized in Triton X-100 suggested that vesicleassociated membrane protein 7 (VAMP7), apolipoprotein B48 (apoB48), liver fatty acid-binding protein (L-FABP), CD36, and the COPII proteins were associated on incubation of the ER with cytosol and ATP. It was proposed that changing cage protein angles could enlarge these protein vesicles to accommodate up to 100-nm-sized cargo (2), this still would not be large enough to transport chylomicrons whose diameter averages 250 nm These considerations led to the proposal that alternative budding machinery was needed for the prechylomicron transport vesicle (PCTV) whose diameter is 350–500 nm (6).
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
