Proteomic analysis of endocytic vesicles

Rab1a regulates motility of early endocytic vesicles

Aparna Mukhopadhyay, Edward Nieves, Fa Yun Che, Jean Wang, Lianji Jin, John W. Murray, Kristie Gordon, Ruth Hogue Angeletti, Allan W. Wolkoff

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Texas-Red-asialoorosomucoid (ASOR) fluorescence-sorted early and late endocytic vesicles from rat liver were subjected to proteomic analysis with the aim of identifying functionally important proteins. Several Rab GTPases, including Rab1a, were found. The present study immunolocalized Rab1a to early and late endocytic vesicles and examined its potential role in endocytosis. Huh7 cells with stable knockdown of Rab1a exhibited reduced endocytic processing of ASOR. This correlated with the finding that Rab1a antibody reduced microtubule-based motility of rat-liver-derived early but not late endocytic vesicles in vitro. The inhibitory effect of Rab1a antibody was observed to be specifically towards minus-end-directed motility. Total and minus-end-directed motility was also reduced in early endocytic vesicles prepared from Rab1a-knockdown cells. These results corresponded with virtual absence of the minus-end-directed kinesin Kifc1 from early endocytic vesicles in Rab1a knockdown cells and imply that Rab1a regulates minus-end-directed motility largely by recruiting Kifc1 to early endocytic vesicles.

Original languageEnglish (US)
Pages (from-to)765-775
Number of pages11
JournalJournal of Cell Science
Volume124
Issue number5
DOIs
StatePublished - Mar 1 2011

Fingerprint

Transport Vesicles
Proteomics
rab GTP-Binding Proteins
Kinesin
Antibodies
Liver
Endocytosis
Microtubules
Fluorescence
Proteins

Keywords

  • Endocytosis
  • Proteomics
  • Rab1a

ASJC Scopus subject areas

  • Cell Biology

Cite this

Proteomic analysis of endocytic vesicles : Rab1a regulates motility of early endocytic vesicles. / Mukhopadhyay, Aparna; Nieves, Edward; Che, Fa Yun; Wang, Jean; Jin, Lianji; Murray, John W.; Gordon, Kristie; Angeletti, Ruth Hogue; Wolkoff, Allan W.

In: Journal of Cell Science, Vol. 124, No. 5, 01.03.2011, p. 765-775.

Research output: Contribution to journalArticle

Mukhopadhyay, Aparna ; Nieves, Edward ; Che, Fa Yun ; Wang, Jean ; Jin, Lianji ; Murray, John W. ; Gordon, Kristie ; Angeletti, Ruth Hogue ; Wolkoff, Allan W. / Proteomic analysis of endocytic vesicles : Rab1a regulates motility of early endocytic vesicles. In: Journal of Cell Science. 2011 ; Vol. 124, No. 5. pp. 765-775.
@article{1b3af5e956a2455ca1940e12de34e28d,
title = "Proteomic analysis of endocytic vesicles: Rab1a regulates motility of early endocytic vesicles",
abstract = "Texas-Red-asialoorosomucoid (ASOR) fluorescence-sorted early and late endocytic vesicles from rat liver were subjected to proteomic analysis with the aim of identifying functionally important proteins. Several Rab GTPases, including Rab1a, were found. The present study immunolocalized Rab1a to early and late endocytic vesicles and examined its potential role in endocytosis. Huh7 cells with stable knockdown of Rab1a exhibited reduced endocytic processing of ASOR. This correlated with the finding that Rab1a antibody reduced microtubule-based motility of rat-liver-derived early but not late endocytic vesicles in vitro. The inhibitory effect of Rab1a antibody was observed to be specifically towards minus-end-directed motility. Total and minus-end-directed motility was also reduced in early endocytic vesicles prepared from Rab1a-knockdown cells. These results corresponded with virtual absence of the minus-end-directed kinesin Kifc1 from early endocytic vesicles in Rab1a knockdown cells and imply that Rab1a regulates minus-end-directed motility largely by recruiting Kifc1 to early endocytic vesicles.",
keywords = "Endocytosis, Proteomics, Rab1a",
author = "Aparna Mukhopadhyay and Edward Nieves and Che, {Fa Yun} and Jean Wang and Lianji Jin and Murray, {John W.} and Kristie Gordon and Angeletti, {Ruth Hogue} and Wolkoff, {Allan W.}",
year = "2011",
month = "3",
day = "1",
doi = "10.1242/jcs.079020",
language = "English (US)",
volume = "124",
pages = "765--775",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "5",

}

TY - JOUR

T1 - Proteomic analysis of endocytic vesicles

T2 - Rab1a regulates motility of early endocytic vesicles

AU - Mukhopadhyay, Aparna

AU - Nieves, Edward

AU - Che, Fa Yun

AU - Wang, Jean

AU - Jin, Lianji

AU - Murray, John W.

AU - Gordon, Kristie

AU - Angeletti, Ruth Hogue

AU - Wolkoff, Allan W.

PY - 2011/3/1

Y1 - 2011/3/1

N2 - Texas-Red-asialoorosomucoid (ASOR) fluorescence-sorted early and late endocytic vesicles from rat liver were subjected to proteomic analysis with the aim of identifying functionally important proteins. Several Rab GTPases, including Rab1a, were found. The present study immunolocalized Rab1a to early and late endocytic vesicles and examined its potential role in endocytosis. Huh7 cells with stable knockdown of Rab1a exhibited reduced endocytic processing of ASOR. This correlated with the finding that Rab1a antibody reduced microtubule-based motility of rat-liver-derived early but not late endocytic vesicles in vitro. The inhibitory effect of Rab1a antibody was observed to be specifically towards minus-end-directed motility. Total and minus-end-directed motility was also reduced in early endocytic vesicles prepared from Rab1a-knockdown cells. These results corresponded with virtual absence of the minus-end-directed kinesin Kifc1 from early endocytic vesicles in Rab1a knockdown cells and imply that Rab1a regulates minus-end-directed motility largely by recruiting Kifc1 to early endocytic vesicles.

AB - Texas-Red-asialoorosomucoid (ASOR) fluorescence-sorted early and late endocytic vesicles from rat liver were subjected to proteomic analysis with the aim of identifying functionally important proteins. Several Rab GTPases, including Rab1a, were found. The present study immunolocalized Rab1a to early and late endocytic vesicles and examined its potential role in endocytosis. Huh7 cells with stable knockdown of Rab1a exhibited reduced endocytic processing of ASOR. This correlated with the finding that Rab1a antibody reduced microtubule-based motility of rat-liver-derived early but not late endocytic vesicles in vitro. The inhibitory effect of Rab1a antibody was observed to be specifically towards minus-end-directed motility. Total and minus-end-directed motility was also reduced in early endocytic vesicles prepared from Rab1a-knockdown cells. These results corresponded with virtual absence of the minus-end-directed kinesin Kifc1 from early endocytic vesicles in Rab1a knockdown cells and imply that Rab1a regulates minus-end-directed motility largely by recruiting Kifc1 to early endocytic vesicles.

KW - Endocytosis

KW - Proteomics

KW - Rab1a

UR - http://www.scopus.com/inward/record.url?scp=79952794950&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79952794950&partnerID=8YFLogxK

U2 - 10.1242/jcs.079020

DO - 10.1242/jcs.079020

M3 - Article

VL - 124

SP - 765

EP - 775

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 5

ER -