Identification of Ser‐1275 and Ser‐1309 as autophosphorylation sites of the insulin receptor

FEBS Letters

Koenen, Marlis³  Meyer, Helmut E⁴  Al-Hasani, Hadi³  Klein, Helmut W²  Müller-Wieland, Dirk¹  Tennagels, Norbert²  Eisermann, Bernd⁴  Magg, Claudia²  Paßlack, Waltraud³ 

[1] Klinik II und Poliklinik für Innere Medizin, Universität zu Köln, Joseph Stelzmannstr. 9, D-50924 Köln, Germany;Institut für Biochemie, Universität zu Köln, Otto-Fischer-Strasse 12–14, D-50674 Köln, Germany;Diabetes Forschungsinstitut Düsseldorf, Auf'm Hennekamp 65, D-40225 Düsseldorf, Germany;Institut für Physiologische Chemie, Ruhr-Universität, Universitätsstrasse 150, D-44780 Bochum, Germany

关键词: Insulin receptor kinase;    Serine autophosphorylation;    Baculovirus expression system;    Kinase negative mutant;    PTH;    phenylthiohydantoin;    SDS-PAGE;    sodium dodecyl sulfate-polyacrylamide electrophoresis;    pY;    phosphotyrosine;    pS;    phosphoserine;    pT;    phosphothreonine;   

DOI  :  10.1016/S0014-5793(96)01342-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have identified Ser-1275 and Ser-1309 as novel serine autophosphorylation sites by direct sequencing of HPLC-purified tryptic phosphopeptides of the histidine-tagged insulin receptor kinase IRKD-HIS. The corresponding peptides (Ser-1275, amino acids 1272–1292; Ser-1309, amino acids 1305–1313) have been detected in the HPLC profiles of both the soluble kinase IRKD, which contains the entire cytoplasmic domain of the insulin receptor β-subunit, and the insulin receptor purified from human placenta. In contrast, a kinase negative mutant, IRKD-K1018A, did not undergo phosphorylation at either the tyrosine or serine residues, strongly suggesting that insulin receptor kinase has an intrinsic activity to autophosphorylate serine residues.

【 授权许可】

Unknown