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Sylvia Müller, Antje Adomeit, Roland Kaufmann, Heribert Appelhans, Hermann Passow, Siegmund Reißmann, Claus Liebmann

Expression and Functional Characterization of a pHis-Tagged Human Bradykinin B2 Receptor in COS-7Cells

A polyHistagged bradykinin (BK) B[2] receptor (pHis BKR) cDNA was constructed and expressed in COS-7 cells. The pHisBKR is suitable for both immunoprecipitation and immunoblotting with antipolyHis antibodies and can be easily purified using NiNTA columns. Immunochemical detection revealed a molecular mass of approximately 66 kDa. The pHisBKR is capable of mediating BKinduced stimulation of inositol phosphate formation as well as of mitogenactivated protein kinase (MAPK) activity. Compared with the wildtype receptor (WTBKR) the tagged receptor showed a slightly enhanced affinity towards BK but a reduced expression level. Despite these modified pharmacological properties the pHistagged BKR may be a useful tool for studying BKR modifications and signaling.

Biological Chemistry, Walter de Gruyter

Print ISSN: 1431-6730
Volume: 381, 04/2000
Pages: 343 - 347

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