Purification and characterization of rabbit muscle acylphosphatase in the thiol (-SH) form.
Modifications in the muscle acylphosphatase purification procedure enabled us to isolate the enzyme with its sole cysteine in the -SH form; this enzyme form is the most abundant in vivo. Our data demonstrates that the enzyme forms purified by previously reported procedures can be easily derived from a reaction of the SH-enzyme with oxidized glutathione. Probably most, or even all, of these enzyme forms are artifacts due to the purification. The SH-acylphosphatase shows kinetic parameters similar to those reported for the mixed disulfide with glutathione and S-S dimer, except for the specific activity value, which is about twice as much, and the Km, which is reduced.
D017766:Acid Anhydride Hydrolases; D000818:Animals; D002846:Chromatography, Affinity; D004220:Disulfides / Q000032:analysis; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification* / Q000378:metabolism; D011817:Rabbits; D013438:Sulfhydryl Compounds; D000097024:Acylphosphatase
D017766:Acid Anhydride Hydrolases; D004220:Disulfides; D010744:Phosphoric Monoester Hydrolases; D013438:Sulfhydryl Compounds; D000097024:Acylphosphatase