Prostaglandin H synthase. Inactivation of the enzyme in the course of catalysis is accompanied by fast and dramatic changes in protein structure.
Prostaglandin H synthase (PGHS) as apo-PGHS, holo-PGHS, and holo-PGHS, inactivated in the course of catalysis was studied using chemical modification with diethyl pyrocarbonate (DEPC). The exhausted reaction with DEPC corresponded to the modification of 7 histidine residues in apo-PGHS and 4 in holo-PGHS. All 18 histidine residues became accessible for modification with DEPC in the enzyme, inactivated in the course of catalysis. The velocities of tryptic cleavage of all the three forms into two fragments were fairly different but independent of modification. Based on the results we hypothesize fast and dramatic changes in the protein structure in the course of the substrate conversion.
Mevkh
A T
AT
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russian Federation.
Miroshnikov
K A
KA
Igumnova
N D
ND
Varfolomeev
S D
SD
D000818:Animals; D001051:Apoenzymes / Q000302:isolation & purification / Q000378:metabolism; D001665:Binding Sites; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology*; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology*; D006639:Histidine; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D011451:Prostaglandin-Endoperoxide Synthases / Q000737:chemistry* / Q000378:metabolism*; D012669:Seminal Vesicles / Q000201:enzymology; D012756:Sheep; D014357:Trypsin
D001051:Apoenzymes; D016861:Cyclooxygenase Inhibitors; D010446:Peptide Fragments; D006639:Histidine; D011451:Prostaglandin-Endoperoxide Synthases; D014357:Trypsin; D004047:Diethyl Pyrocarbonate