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Regulation of glycolytic enzyme activity during chronic hypoxia by changes in rate limiting enzyme content use of mono clonal antibodies to quantitate changes in pyruvate kinase ec content

, : Regulation of glycolytic enzyme activity during chronic hypoxia by changes in rate limiting enzyme content use of mono clonal antibodies to quantitate changes in pyruvate kinase ec content. Journal of Clinical Investigation 66(6): 1258-1264

Monoclonal antibodies were prepared against pyruvate kinase (PyKi; EC and used to quantitate PyKi content in L2 lung cells and [human female embryo lung] WI-38 fibroblasts cultivated under hypoxic and normoxic conditions. After 96 h of hypoxic cultivation, PyKi activity was significantly increased in both cell types (L2: normoxia [PO2 = 142 torr], 0.11 .+-. 0.01 [SD]; hypoxia [PO2 = 14 torr], 0.25 .+-. 0.04 U/.mu.g DNA, P < 0.01). PyKi content increased proportionately in both cell lines (L2: normoxia, 0.44 .+-. 0.13; hypoxia, 0.94 .+-. 0.13 .mu.g enzyme protein/.mu.g DNA). Specific activity was not significantly different after 96 h (L2: normoxia, 261 .+-. 11; hypoxia, 261 .+-. 14 U/mg enzyme protein). Regulation of glycolysis during chronic hypoxia occurs at the level of enzyme content. Chronic O2 depletion leads to either an increased rate of biosynthesis or a decreased rate of biodegradation of PyKi, causing augmented glycolytic capacity. Monoclonal antibodies provide a highly specific, convenient approach to characterizing enzymes, as well as quantitating cellular enzyme content.

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