Quantitative analysis of liver Golgi proteome in the cell cycle.
During mitosis, the Golgi membranes in mammalian cells undergo a continuous disassembly process and generate mitotic fragments that are distributed into the daughter cells and reassembled into new Golgi after mitosis. This disassembly and reassembly process is critical for Golgi biogenesis during cell division, but the underlying molecular mechanism is poorly understood. In this study, we have recapitulated this process using an in vitro assay and analyzed the proteins that are associated with interphase and mitotic Golgi membranes using quantitative proteomics that combines the isobaric tags for relative and absolute quantification approach with OFFGEL isoelectric focusing separation and LC-MALDI-MS/MS. A total of 1,193 Golgi-associated proteins were identified and quantified. These included broad functional categories: Golgi structural proteins, Golgi resident enzymes, SNAREs, Rab GTPases, and secretory and cytoskeletal proteins. More importantly, the combination of the quantitative proteomic approach with Western blot analysis allowed us to unveil 86 proteins with significant changes in abundance under the mitotic condition compared to the interphase condition. Altogether, this systematic quantitative proteomic study revealed candidate proteins of the molecular machinery that controls the Golgi disassembly and reassembly processes in the cell cycle.
Department of Physiology, Wayne State University, Detroit, MI, USA. xchen@med.wayne.edu
Xuequn
X
Chen
Philip C
PC
Andrews
Yanzhuang
Y
Wang
D000818:Animals; D002458:Cell Fractionation; D002474:Cell-Free System; D005260:Female; D006056:Golgi Apparatus / Q000737:chemistry / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D007425:Intracellular Membranes / Q000737:chemistry / Q000378:metabolism*; D007525:Isoelectric Focusing; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008938:Mitosis; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000737:chemistry
D008565:Membrane Proteins; D010446:Peptide Fragments; D020543:Proteome; D014357:Trypsin