Volume 1041, Issue 1 p. 332-337

Responses of GPCR135 to Human Gene 3 (H3) Relaxin in CHO-K1 Cells Determined by Microphysiometry

EMMA T. VAN DER WESTHUIZEN

EMMA T. VAN DER WESTHUIZEN

Department of Pharmacology, Monash University, Clayton, Victoria 3168, Australia

Howard Florey Institute, University of Melbourne, Victoria 3010, Australia

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PATRICK M. SEXTON

PATRICK M. SEXTON

Howard Florey Institute, University of Melbourne, Victoria 3010, Australia

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ROSS A. D. BATHGATE

ROSS A. D. BATHGATE

Howard Florey Institute, University of Melbourne, Victoria 3010, Australia

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ROGER J. SUMMERS

Corresponding Author

ROGER J. SUMMERS

Department of Pharmacology, Monash University, Clayton, Victoria 3168, Australia

Address for correspondence: Prof. R. J. Summers, Department of Pharmacology, Monash University, Wellington Road, Clayton 3800, Australia. Voice: +61-3-9905-1440; fax: +61-3-9905-8192. [email protected]Search for more papers by this author
First published: 09 January 2006
Citations: 32

Abstract

Abstract: This study examined the functional response to human relaxin 2 (H2 relaxin), human relaxin 3 (H3 relaxin), porcine relaxin, and human INSL3 in the cytosensor microphysiometer, using CHO-K1 cells stably expressing human GPCR135. CHO-K1 cells stably expressing GPCR135 were generated by the serial dilution method and receptor properties were assessed. Saturation studies of [125I] H3 relaxin binding to GPCR135 in these cells gave a Bmax of 32.61 ± 6.5 fmol/mg protein and Kd of 0.12 ± 0.08 nM. The functional response to H3 relaxin and other relaxin/insulin peptides of GPCR135 expressed in CHO-K1 cells was measured in the cytosensor microphysiometer and analyzed using inhibitors of signal transduction proteins.