Comparative proteomics of clathrin-coated vesicles

• DOI: 10.1083/jcb.200607164
• Publication Date: 2006-11-20T19:51:13Z
• Authors: Georg H.H. Borner; Michael Harbour; Svenja Hester; Kathryn S. Lilley; Margaret S. Robinson
• Keywords: Clathrin adaptor proteins; Organelle; COPI; Proteome

Clathrin-coated vesicles (CCVs) facilitate the transport of cargo between trans-Golgi network, endosomes, and plasma membrane. This study presents first comparative proteomics investigation CCVs. A CCV-enriched fraction was isolated from HeLa cells a “mock CCV” clathrin-depleted cells. We used combination 2D difference gel electrophoresis isobaric tags for relative absolute quantification (iTRAQ) in conjunction with mass spectrometry to analyze compare two fractions. In total, 63 bona fide CCV proteins were identified, including 28 whose association CCVs had not previously been established. These include numerous post-Golgi SNAREs; subunits AP-3, retromer, BLOC-1 complexes; lysosomal enzymes; CHC22; five novel unknown function. The strategy outlined this paper should be widely applicable as means distinguishing genuine organelle components contaminants.