title: Inactivation of muscle adenylate kinase by site-specific destruction of tyrosine 95 using potassium ferrate.
abstract: Potassium ferrate, an analog of orthophosphate and a potent oxidizing agent, was found to irreversibly inactivate porcine muscle adenylate kinase. Inhibition was prevented by competitive inhibitors or substrates, indicating that the action of ferrate was site-specific. Inactivation was accompanied by the loss of Cys-25 and Tyr-95. P1,P5-di(adenosine 5')-pentaphosphate (10(-7) M), a powerful competitive inhibitor, gave 50% protection to the enzyme from ferrate inactivation. No loss of tyrosine or cysteine residues was observed under conditions of total protection. The degree of inactivation was proportional to the amount of Tyr-95 destroyed. However, Cys-25 was totally oxidized when only 55% inactivation had occurred. Partially inactivated enzyme exhibited a Km for ATP and AMP similar to that of the untreated enzyme. It appears that Cys-25 may be proximate to a phosphate-binding site but is not directly involved in the catalytic reaction. The results suggest that Tyr-95 is located in the vicinity of a phosphate-binding region of adenylate kinase and is essential for enzyme activity.
authors.forename: M D
authors.initials: MD
authors.lastname: Crivellone
authors.forename: M
authors.initials: M
authors.lastname: Hermodson
authors.forename: B
authors.initials: B
authors.lastname: Axelrod
mesh_terms: D000263:Adenylate Kinase / Q000037:antagonists & inhibitors*; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003545:Cysteine; D007501:Iron / Q000494:pharmacology*; D058085:Iron Compounds; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D010770:Phosphotransferases / Q000037:antagonists & inhibitors*; D011188:Potassium / Q000494:pharmacology*; D017680:Potassium Compounds; D013329:Structure-Activity Relationship; D013552:Swine; D014443:Tyrosine
chemical_list: D000596:Amino Acids; D058085:Iron Compounds; D017680:Potassium Compounds; C044384:potassium ferrate; D014443:Tyrosine; D007501:Iron; D010770:Phosphotransferases; D000263:Adenylate Kinase; D003545:Cysteine; D011188:Potassium