14-3-3-affinity purification of over 200 human phosphoproteins reveals new links to regulation of cellular metabolism, proliferation and trafficking.

摘要: 14-3-3-interacting proteins were isolated from extracts of proliferating HeLa cells using 14-3-3 affinity chromatography, eluting with a phosphopeptide that competes targets for binding. The did not bind to (14-3-3s) after dephosphorylation protein phosphatase 2A (PP2A), indicating binding 14-3-3s requires their phosphorylation. identified by tryptic mass fingerprinting and Western blotting include many enzymes involved in generating precursors such as purines (AMP, GMP ATP), FAD, NADPH, cysteine S-adenosylmethionine, which are needed cell growth, regulators proliferation, including DNA replication, anti-oxidative metabolism, actin dynamics cellular trafficking, whose deregulation has been implicated cancers, diabetes, Parkinsonism other neurological diseases. Several bound 14-3-3-Sepharose cells, but non-proliferating, serum-starved novel microtubule-interacting ELP95 (EMAP-like 95 kDa) small HVA22/Yop1p-related protein. In contrast, the interactions N-methyl-D-aspartate receptor subunit NuMA (nuclear mitotic apparatus protein) regulated serum. Overall, our findings suggest may be central integrating regulation biosynthetic survival, processes human cells.