Surface plasmon resonance using the catalytic domain of soluble guanylate cyclase allows the detection of enzyme activators.
Soluble Guanylate Cyclase (sGC) is the receptor for the signalling agent nitric oxide (NO) and catalyses the production of the second messenger cyclic guanosine monophosphate (cGMP) from guanosine triphosphate (GTP). The enzyme is an attractive drug target for small molecules that act in the cardiovascular and pulmonary systems, and has also shown to be a potential target in neurological disorders. We have discovered that 5-(indazol-3-yl)-1,2,4-oxadiazoles activate the enzyme in the absence of added NO and shown they bind to the catalytic domain of the enzyme after development of a surface plasmon resonance assay that allows the biophysical detection of intrinsic binding of ligands to the full length sGC and to a construct of the catalytic domain.
The Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, United Kingdom.
Filipa
F
Mota
Structural Genomics Consortium, University of Oxford, United Kingdom.
Charles K
CK
Allerston
The Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, United Kingdom.
Kathryn
K
Hampden-Smith
The Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, United Kingdom.
John
J
Garthwaite
The Wolfson Institute for Biomedical Research, University College London, Gower Street, London WC1E 6BT, United Kingdom.. Electronic address: d.selwood@ucl.ac.uk.
David L
DL
Selwood
D055162:Biocatalysis; D020134:Catalytic Domain / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D006157:Guanosine Monophosphate / Q000096:biosynthesis; D006162:Guanylate Cyclase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015394:Molecular Structure; D010069:Oxadiazoles / Q000737:chemistry / Q000494:pharmacology*; D018160:Receptors, Cytoplasmic and Nuclear / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000071756:Soluble Guanylyl Cyclase; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance
C000592082:5-(indazol-3-yl)-1,2,4-oxadiazole; D010069:Oxadiazoles; D018160:Receptors, Cytoplasmic and Nuclear; D006157:Guanosine Monophosphate; D006162:Guanylate Cyclase; D000071756:Soluble Guanylyl Cyclase
Biophysical techniques; Enzyme activators; Nitric oxide; Soluble guanylate cyclase; Surface plasmon resonance