Using proton nuclear magnetic resonance to study the mode of ribonuclease A inhibition by competitive and noncompetitive inhibitors.
The C(2) proton resonances of the active site histidines (His 12 and His 119) of ribonuclease A have been exploited to study the inhibition pattern of both noncompetitive (four green tea polyphenols and their copper complexes) and competitive (3'-O-carboxy esters of thymidine and 3'-amino derivatives of uridine) inhibitors. Competitive inhibitors devoid of any phosphate group have the ability to change the pK(a) of the histidine residues at the active site. Their mode of inhibition, albeit competitive, is found to be different compared to known phosphate inhibitors 2'-CMP and 3'-CMP as revealed by changes in the pK(a) values. We find a correlation between the changes in the chemical shift of His 12 and the corresponding inhibition constants (K(i)).
Ghosh
Kalyan Sundar
KS
Department of Chemistry, Indian Institute of Technology, Kharagpur, West Bengal 721302, India.
Debnath
Joy
J
Pathak
Tanmaya
T
Dasgupta
Swagata
S
D001667:Binding, Competitive; D020134:Catalytic Domain; D002392:Catechin / Q000737:chemistry; D003300:Copper / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry*; D005419:Flavonoids / Q000737:chemistry; D006639:Histidine / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008956:Models, Chemical; D010636:Phenols / Q000737:chemistry; D059808:Polyphenols; D011522:Protons; D012259:Ribonuclease, Pancreatic / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D013662:Tea; D013936:Thymidine / Q000737:chemistry; D014529:Uridine / Q000737:chemistry
D004791:Enzyme Inhibitors; D005419:Flavonoids; D010636:Phenols; D059808:Polyphenols; D011522:Protons; D013662:Tea; D006639:Histidine; D003300:Copper; D002392:Catechin; D012259:Ribonuclease, Pancreatic; D013936:Thymidine; D014529:Uridine