Two-dimensional mapping of the endogenous proteins of the rat hepatocyte Golgi complex cleared of proteins in transit

Electrophoresis
R S TaylorK E Howell

Abstract

The discovery of additional endogenous Golgi proteins will lead to significant new insights into Golgi function. To this end, stacked Golgi fractions (SGFs) were isolated from rat liver before (CTL SGF) and after molecules in transit through the Golgi were cleared by pre-treatment with cycloheximide (CHX SGF). Electron microscopic (EM) morphometric and biochemical analyses showed that the in vivo stacked morphology is retained, that > 90% of the elements can be positively identified as Golgi stacks and cisternae, and that transmembrane protein markers of the Golgi complex are enriched 300- to 800-fold over starting postnuclear supernatant (PNS). High-resolution two-dimensional (2-D) gel mapping has been carried out on the CTL PNS, CTL SII (an intermediate fraction), CTL SGF, CHX SGF, CHX SGF - high pH supernatant, and CHX SGF - high pH pellet. This analysis, coupled with immunoblotting and alignment of the 2-D gels with master gels, has allowed the identification of a number of known proteins and the preliminary characterization of the most abundant 173 Golgi-specific proteins. These 173 proteins have been placed into three categories: cargo, cytosolic Golgi-associated, and resident Golgi proteins. These categories are tentativ...Continue Reading

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Citations

Nov 18, 2000·Electrophoresis·E JungD F Hochstrasser
May 28, 2003·Mass Spectrometry Reviews·Mathias Dreger
Mar 6, 1999·Journal of Chromatography. B, Biomedical Sciences and Applications·C PasqualiL A Huber
Apr 11, 2013·Chemical Reviews·Chad P SatoriEdgar A Arriaga
Feb 13, 2003·European Journal of Biochemistry·Mathias Dreger
Nov 18, 2003·The Plant Journal : for Cell and Molecular Biology·Min Seok BaeOhkmae K Park
Apr 29, 2000·Electrophoresis·G L CorthalsJ C Sanchez
May 17, 2003·Circulation Research·Lukas A HuberIlja Vietor
Feb 24, 2001·Traffic·W H McDonald, J R Yates
Jun 21, 2007·Journal of Proteome Research·Giuseppe Gazzana, Jürgen Borlak

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