Purification, characterization and inhibition by MK-401 of Fasciola hepatica phosphoglyceromutase

• Published in: Molecular and biochemical parasitology Vol. 5; no. 5; pp. 321 - 332
• Main Authors: Schulman, Marvin D., Valentino, Delia
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.1982
• Subjects: 2,3-Diphosphoglycerate; Animals; Chloromercuribenzoates - pharmacology; Diphosphoglyceric Acids - pharmacology; drugs; enzymes; Ethylmaleimide - pharmacology; Fasciola hepatica; Fasciola hepatica - enzymology; Iodoacetamide - pharmacology; Kinetics; Liver fluke; mechanism of action; MK-401; Molecular Weight; Phosphoglycerate Mutase - antagonists & inhibitors; Phosphoglycerate Mutase - isolation & purification; Phosphoglycerate Mutase - metabolism; Phosphoglyceromutase; Phosphotransferases - metabolism; purification; Trematoda; SDS; DTT; EtSH; IAA; EDTA; PCMB; Fasciola hepatica; 3-PGA; Liver fluke; NEM; DNTB; 2,3-DiPGA; Phosphoglyceromutase; MK-401
• ISSN: 0166-6851; 1872-9428
• DOI: 10.1016/0166-6851(82)90039-1

Phosphoglyceromutase (EC 2.7.5.3) of Fasciola hepatica was purified 1390-fold to homogeneity. The enzyme had a molecular weight of 120 000 and was a tetramer composed of identical 30 000 molecular weight subunits. The enzyme was 2,3-diphosphoglyceric acid dependent, possessed reactive sulfhydryl groups and was inhibited irreversibly by iodoacetamide, and N-ethylmaleimide and reversibly by p-chloromercuribenzoate and 5,5′-dithiobis(2-nitrobenzoic acid). Initial velocity studies suggest that reaction occurred via a sequential mechanism and that MK-401 was a competitive inhibitor versus both 3-phosphoglyceric acid and 2,3-diphosphoglyceric acid.