Abstract

The effect of hypoxia on the levels of glycogen, glucose and lactate as well as the activities and binding of glycolytic and associated enzymes to subcellular structures was studied in brain, liver and white muscle of the teleost fish, Scorpaena porcus. Hypoxia exposure decreased glucose levels in liver from 2.53 to 1.70 mumol/g wet weight and in muscle led to its increase from 3.64 to 25.1 mumol/g wet weight. Maximal activities of several enzymes in brain were increased by hypoxia: hexokinase by 23%, phosphoglucoisomerase by 47% and phosphofructokinase (PFK) by 56%. However, activities of other enzymes in brain as well as enzymes in liver and white muscle were largely unchanged or decreased during experimental hypoxia. Glycolytic enzymes in all three tissues were partitioned between soluble and particulate-bound forms. In several cases, the percentage of bound enzymes was reduced during hypoxia; bound aldolase in brain was reduced from 36.4 to 30.3% whereas glucose-6-phosphate dehydrogenase fell from 55.7 to 28.7% bound. In muscle PFK was reduced from 57.4 to 41.7% bound. Oppositely, the proportion of bound aldolase and triosephosphate isomerase increased in hypoxic muscle. Phosphoglucomutase did not appear to occur in a bound form in liver and bound phosphoglucomutase disappeared in muscle during hypoxia exposure. Anoxia exposure also led to the disappearance of bound fructose-1,6-bisphosphatase in liver, whereas a bound fraction of this enzyme appeared in white muscle of anoxic animals. The possible function of reversible binding of glycolytic enzymes to subcellular structures as a regulatory mechanism of carbohydrate metabolism is discussed.

Highlights

  • Glycolytic and associated enzymes can bind reversibly to various cellular structures such as glycogen particles, F-actin, microfilaments [1,2,3,4,5] and membranes [6,7,8]

  • Exposure of fish to water with 15% oxygen saturation for 90 min did not significantly change the glycogen content of any tissue, a definite trend to reduced mean glycogen levels after hypoxia was observed in all instances

  • Glucose was unaltered after hypoxia exposure whereas in hypoxic liver glucose decreased by 33%

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Summary

Introduction

Glycolytic and associated enzymes can bind reversibly to various cellular structures such as glycogen particles, F-actin, microfilaments [1,2,3,4,5] and membranes [6,7,8]. These interactions can change some of their functional properties. Catalytic properties can be modified by enzyme interaction with structural elements [1,2,7,10,11]. Different attempts have been made to stabilize such a complex such as the addition of crowding agents to the extraction medium (e.g. polyethylene glycol) that should increase the stability of enzyme-enzyme interactions during their isolation [14,15]

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