Proteomic Analysis of the Very Low Density Lipoprotein (VLDL) transport vesicles

Abdul Rahim, Erika Nafi-valencia, Shaila Siddiqi, Riyaz Basha, Chukwuemeka C. Runyon, Shadab A. Siddiqi

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Abstract

The VLDL transport vesicle (VTV) mediates the transport of nascent VLDL particles from the ER to the Golgi and plays a key role in VLDL-secretion from the liver. The functionality of VTV is controlled by specific proteins; however, full characterization and proteomic profiling of VTV remain to be carried out. Here, we report the first proteomic profile of VTVs. VTVs were purified to their homogeneity and characterized biochemically and morphologically. Thin section transmission electron microscopy suggests that the size of VTV ranges between 100. nm to 120. nm and each vesicle contains only one VLDL particle. Immunoblotting data indicate VTV concentrate apoB100, apoB48 and apoAIV but exclude apoAI. Proteomic analysis based on 2D-gel coupled with MALDI-TOF identified a number of vesicle-related proteins, however, many important VTV proteins could only be identified using LC-MS/MS methodology. Our data strongly indicate that VTVs greatly differ in their proteome with their counterparts of intestinal origin, the PCTVs. For example, VTV contains Sec22b, SVIP, ApoC-I, reticulon 3, cideB, LPCAT3 etc. which are not present in PCTV. The VTV proteome reported here will provide a basic tool to study the mechanisms underlying VLDL biogenesis, maturation, intracellular trafficking and secretion from the liver.

Original languageEnglish
Pages (from-to)2225-2235
Number of pages11
JournalJournal of Proteomics
Volume75
Issue number7
DOIs
StatePublished - 3 Apr 2012

Keywords

  • Apolipoprotein B (apoB)
  • Endoplasmic reticulum (ER)
  • Triacylglycerol (TAG)
  • VLDL transport vesicle (VTV)
  • Very Low-Density Lipoprotein (VLDL)

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    Rahim, A., Nafi-valencia, E., Siddiqi, S., Basha, R., Runyon, C. C., & Siddiqi, S. A. (2012). Proteomic Analysis of the Very Low Density Lipoprotein (VLDL) transport vesicles. Journal of Proteomics, 75(7), 2225-2235. https://doi.org/10.1016/j.jprot.2012.01.026