TY - JOUR
T1 - Proteomic Analysis of the Very Low Density Lipoprotein (VLDL) transport vesicles
AU - Rahim, Abdul
AU - Nafi-valencia, Erika
AU - Siddiqi, Shaila
AU - Basha, Riyaz
AU - Runyon, Chukwuemeka C.
AU - Siddiqi, Shadab A.
N1 - Funding Information:
This study was supported by NIH 's DK-81413 (to SAS) from the National Institute of Diabetes And Digestive and Kidney Diseases . The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Diabetes And Digestive and Kidney Diseases or the National Institutes of Health.
PY - 2012/4/3
Y1 - 2012/4/3
N2 - 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.
AB - 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.
KW - Apolipoprotein B (apoB)
KW - Endoplasmic reticulum (ER)
KW - Triacylglycerol (TAG)
KW - VLDL transport vesicle (VTV)
KW - Very Low-Density Lipoprotein (VLDL)
UR - http://www.scopus.com/inward/record.url?scp=84858751786&partnerID=8YFLogxK
U2 - 10.1016/j.jprot.2012.01.026
DO - 10.1016/j.jprot.2012.01.026
M3 - Article
C2 - 22449872
AN - SCOPUS:84858751786
SN - 1874-3919
VL - 75
SP - 2225
EP - 2235
JO - Journal of Proteomics
JF - Journal of Proteomics
IS - 7
ER -