Identification of protein tyrosine phosphatase 1B and casein as substrates for 124-v-Mos
Proikas-Cezanne, Tassula, Stabel, Silvia and Riethmacher, Dieter (2002) Identification of protein tyrosine phosphatase 1B and casein as substrates for 124-v-Mos. BMC Biochemistry, 3, (6), 1-7. (doi:10.1186/1471-2091-3-6).
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The mos proto-oncogene encodes a cytoplasmic serine/threonine-specific protein kinase with crucial function during meiotic cell division in vertebrates. Based on oncogenic amino acid substitutions the viral derivative, 124-v-Mos, displays constitutive protein kinase activity and functions independent of unknown upstream effectors of mos protein kinase. We have utilized this property of 124-v-Mos and screened for novel mos substrates in immunocomplex kinase assays in vitro.
We generated recombinant 124-v-Mos using the baculovirus expression system in Spodoptera frugiperda cells and demonstrated constitutive kinase activity by the ability of 124-v-Mos to auto-phosphorylate and to phosphorylate vimentin, a known substrate of c-Mos. Using this approach we analyzed a panel of acidic and basic substrates in immunocomplex protein kinase assays and identified novel in vitro substrates for 124-v-Mos, the protein tyrosine phosphatase 1B (PTP1B), alpha-casein and beta-casein. We controlled mos-specific phosphorylation of PTP1B and casein in comparative assays using a synthetic kinase-inactive 124-v-Mos mutant and further, tryptic digests of mos-phosphorylated beta-casein identified a phosphopeptide specifically targeted by wild-type 124-v-Mos. Two-dimensional phosphoamino acid analyses showed that 124-v-mos targets serine and threonine residues for phosphorylation in casein at a 1:1 ratio but auto-phosphorylation occurs predominantly on serine residues.
The mos substrates identified in this study represent a basis to approach the identification of the mos-consensus phosphorylation motif, important for the development of specific inhibitors of the Mos protein kinase.
|Digital Object Identifier (DOI):||doi:10.1186/1471-2091-3-6|
|Keywords:||cell line,non-receptor type 1, caseins, in vitro, proto-oncogene proteins c-mos, threonine, phosphorylation, protein-serine-threonine kinases, protein tyrosine phosphatase, genetics|
R Medicine > RM Therapeutics. Pharmacology
Q Science > QR Microbiology
|Divisions:||University Structure - Pre August 2011 > School of Medicine > Infection, Inflammation and Repair
University Structure - Pre August 2011 > School of Medicine
University Structure - Pre August 2011 > School of Medicine > Human Genetics
|Date Deposited:||08 Sep 2008|
|Last Modified:||31 Mar 2016 12:42|
|RDF:||RDF+N-Triples, RDF+N3, RDF+XML, Browse.|
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