Calmodulin interaction with peptides from G-protein coupled receptors measured with S-Tag labeling |
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Authors: | Zhang Ying Wang Danxin Sadée Wolfgang |
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Institution: | Department of Pharmacology, College of Medicine and Public Health, Ohio State University, Columbus, OH 43210, USA. |
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Abstract: | We have developed a quantitative assay of calmodulin (CaM) binding to S-Tag labeled peptides derived from G-protein coupled receptor (GPCR) sequences. CaM binding of peptides derived from the third intracellular loop (i3) of mu opioid receptor (MOR) was confirmed and the CaM-binding motif refined. A MORi3 peptide with a Lys > Ala substitution--shown to reduce CaM-binding of intact MOR--bound fivefold less avidly than the wild-type peptide. Screening peptides derived from i3 loops of other GPCR families confirmed 5HT1A, and identified muscarinic receptor 3, and melanocortin receptor 1, as proteins carrying CaM-binding domains. The use of S-Tag labeling can serve for rapid screening of putative CaM-binding domains in GPCRs. |
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Keywords: | Calmodulin G-protein coupled receptors In vitro translation μ opioid receptor |
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