Human recombinant soluble guanylyl cyclase: Expression, purification, and regulation
نویسندگان
چکیده
منابع مشابه
Human recombinant soluble guanylyl cyclase: expression, purification, and regulation.
The alpha1- and beta1-subunits of human soluble guanylate cyclase (sGC) were coexpressed in the Sf9 cells/baculovirus system. In addition to the native enzyme, constructs with hexahistidine tag at the amino and carboxyl termini of each subunit were coexpressed. This permitted the rapid and efficient purification of active recombinant enzyme on a nickel-affinity column. The enzyme has one heme p...
متن کاملNucleotidyl cyclase activity of recombinant soluble guanylyl cyclase
Background The ubiquitously expressed soluble guanylyl cyclase (sGC) converts guanosine 5'-triphosphate (GTP) to guanosine 3':5'-cyclic monophosphate (cGMP). The heterodimeric protein is activated by nitric oxide (NO). sGC plays a key role in the regulation of vascular tone and neurotransmission. Hence, sGC is an important target for the treatment of cardiovascular diseases e.g. pulmonary hyper...
متن کاملRegulation of soluble guanylyl cyclase by phosphorylation
Results In vitro kinase assays revealed that the α1, but not the β1, subunit of sGC is a PKG substrate and that the phosphorylation site is located within the first 360 residues of the α1. A constitutively active form of PKG stimulated incorporation of 32P into the α1 subunit in vivo. In addition, PKG could be detected in sGC immunoprecipitates, suggesting that the two proteins interact in cell...
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Betatrophin is a member of the angiopoietin-like (ANGPTL) family that has been implicated in both triglyceride and glucose metabolism. The physiological functions and molecular targets of this protein remain largely unknown; hence, a purified available protein would aid study of the exact role of betatrophin in lipid or glucose metabolism. In this study, we cloned the full-length cDNA of betatr...
متن کاملRegulation of soluble guanylyl cyclase: looking beyond NO.
NEUROTRANSMISSION, PLATELET FUNCTION, skeletal muscle and vascular and nonvascular smooth muscle, and numerous other cellular functions are significantly regulated by the principal enzyme initially transducing nitric oxide (NO) signals, soluble guanylyl cyclase (sGC) (8). The better-characterized active forms of this enzyme exist as a heterodimer, comprising an and a heme-containing -sGC subuni...
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 2000
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.190333697