METHODS: We treated 3T3 - L1 adipocytes with 2.5 mmol / l R (+) alpha - lipoic acid for 2 to 60 min, followed by assays of: 2 - deoxyglucose uptake; glucose transporter 1 and 4 (GLUT1 and GLUT4) subcellular localization; tyrosine phosphorylation of the insulin receptor or of the insulin receptor substrate - 1 in cell lysates; association of phosphatidylinositol 3 - kinase activity with immunoprecipitates of proteins containing phosphotyrosine or of insulin receptor substrate - 1 using a in vitro kinase assay; association of the p85 subunit of phosphatidylinositol 3 - kinase with
phosphotyrosine proteins or with insulin receptor substrate - 1; and in vitro activity of immunoprecipitated Akt1.
«Yingchun has developed methods to analyze both low and high abundant
phosphotyrosine proteins from the pseudopodia,» says Klemke.
Not exact matches
After extracting all the
proteins they could get, Hunter and his colleagues would measure the amount of
phosphotyrosine.
For example, the insulin receptor substrate and
phosphotyrosine binding
proteins GAB1 / GAB2 emerged on the metazoan stem after the divergence of choanoflagellates, indicating that an insulin - signalling - like pathway may have been a key regulator of growth in early animals by tying into the ancient PDK1 and Akt kinases (Fig. 2b).
STAT1 and STAT2 represent two of the seven mammalian STAT
proteins that share several structurally and functionally conserved domains, including an NH2 - terminal coiled - coil region for interactions with other transcription factors and chaperone
proteins, a
phosphotyrosine - binding Src homology 2 domain required for receptor binding and STAT dimerization, and a COOH - terminal transcription activation domain (TAD) within which a highly conserved tyrosine residue is present.