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Enzyme inactivation by a cellular neutral protease: Enzyme specificity, effects of ligands on inactivation, and implications for the regulation of enzyme degradation
Author(s) -
Levy Michael R.,
McConkey Charles L.
Publication year - 1977
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.1040900211
Subject(s) - biochemistry , enzyme , protease , malate dehydrogenase , dehydrogenase , fumarase , isocitrate dehydrogenase , tmprss6 , biology , citric acid cycle , chemistry , serine protease
A protease from Tetrahymena pyriformis inactivated eight of nine commercially available enzymes tested, including lactate dehydrogenase, isocitrate dehydrogenase (TPN‐specific), glucose‐6 phosphate dehydrogenase, D‐amino acid oxidase, fumarase, pyruvate kinase, hexokinase, and citrate synthase. Urate oxidase was not inactivated. Inactivation occurred at neutral pH, was prevented by inhibitors of the protease, and followed first order kinetics. In those cases tested, inactivation was enhanced by mercaptoethanol. Most of the enzyme‐inactivating activity was due to a protease of molecular weight 25,000 that eluted from DEAE‐Sephadex at 0.3 M KCl. A second protease of this molecular weight, which was not retained by the gel, inactivated only isocitrate dehydrogenase and D‐amino acid oxidase. These two proteases could also be distinguished by temperature and inhibitor sensitivity. Two other protease peaks obtained by DEAE‐Sephadex chromatography had little or no enzyme inactivating activity, while another attacked only D‐amino acid oxidase. At least six of the enzymes could be protected from proteolytic inactivation by various ligands. Isocitrate dehydrogenase was protected by isocitrate, TPN, or TPNH, glucose‐6‐dehydrogenase by glucose‐6‐P or TPN, pyruvate kinase by phosphoenolypyruvate or ADP, hexokinase by glucose, and fumarase by a mixture of fumarate and malate. Lactate dehydrogenase was not protected by either of its substrates or coenzymes. Citrate synthase was probably protected by oxalacetate. Our data suggest that the protease or proteases discussed here may participate in the inactivation or degradation of at least some enzymes in Tetrahymena . Since the inactivation occurs at neutral pH, this process could be regulated by variations in the cellular levels of substrates, coenzymes, or allosteric regulators resulting from changes in growth conditions or growth state. Such a mechanism would permit the selective retention of enzymes of metabolically active pathways.