I've tried numerous times to express a receptor guanylyl cyclase in mammalian cells, only to find that the protein is constitutively active (verified by assaying for cGMP, the product of its catalyzed reaction). I've tried two different cell lines (HEK293T, CHO), made sure the media was free of any potential ligands for the receptor, etc., but I'm a little at a loss for what else I should be looking at. This protein has been expressed similarly as reported in the literature, with no constitutive activity. Anyone have any ideas, or encountered something similar? Thanks!
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Transfected protein is constitutively active
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