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Enzyme activity profiles in an overwintering population of freeze-tolerant larvae of the gall fly, Eurosta solidaginis

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Abstract

The activity of some enzymes of intermediary metabolism, including enzymes of glycolysis, the hexose monophosphate shunt, and polyol cryoprotectant synthesis, were measured in freeze-tolerant Eurosta solidaginis larvae over a winter season and upon entry into pupation. Flexible metabolic rearrangement was observed concurrently with acclimatization and development. Profiles of enzyme activities related to the metabolism of the cryoprotectant glycerol indicated that fall biosynthesis may occur from two possible pathways: 1. glyceraldehyde-phosphate → glyceraldehyde → glycerol, using glyceraldehyde phosphatase and NADPH-linked polyol dehydrogenase, or 2. dihydroxyacetonephosphate → glycerol-3-phosphate → glycerol, using glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase. Clearance of glycerol in the spring appeared to occur by a novel route through the action of polyol dehydrogenase and glyceraldehyde kinase. Profiles of enzyme activities associated with sorbitol metabolism suggested that this polyol cryoprotectant was synthesized from glucose-6-phosphate through the action of glucose-6-phosphatase and NADPH-linked polyol dehydrogenase. Removal of sorbitol in the spring appeared to occur through the action of sorbitol dehydrogenase and hexokinase. Glycogen phosphorylase activation ensured the required flow of carbon into the synthesis of both glycerol and sorbitol. Little change was seen in the activity of glycolytic or hexose monophosphate shunt enzymes over the winter. Increased activity of the α-glycerophosphate shuttle in the spring, indicated by greatly increased glycerol-3-phosphate dehydrogenase activity, may be key to removal and oxidation of reducing equivalents generated from polyol cryoprotectan catabolism.

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Abbreviations

6PGDH:

6-Phosphogluconate dehydrogenase

DHAP:

dihydroxy acetone phosphate

F6P:

fructose-6-phosphate

F6Pase:

fructose-6-phospha-tase

FBPase:

fructose-bisphosphatase

G3P:

glycerol-3-phosphate

G3Pase:

glycerol-3-phosphate phophatase

G3PDH:

glycerol-3-phosphate dehydrogenase

G6P:

glucose-6-phosphate

G6Pase:

glucose-6-phosphatase

G6PDH:

glucose-6-phosphate dehydrogenase

GAK:

glyceraldehyde kinase

GAP:

glyceraldehyde-3-phosphate

GAPase:

glyceraldehyde-3-phosphatase

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

GDH:

glycerol dehydrogenase

GPase:

glycogen phosphorylase

HMS:

hexose monophosphate shunt

LDH:

lactate dehydrogenase

NADP-IDH:

NADP+-dependent isocitrate dehydrogenase

PDHald:

polyol dehydrogenase, glyceraldehyde activity

PDHgluc:

polyol dehydrogenase, glucose activity

PFK:

phosphofructokinase

PGI:

phosphoglucoisomerase

PGK:

phosphoglycerate kinase

PGM:

phosphoglucomutase

PK:

pyruvate kinase

PMSF:

phenylmethylsulfonylfluoride

SoDH:

sorbitol dehydrogenase

V max :

maximal enzyme activity

ww:

wet weight

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Joanisse, D.R., Storey, K.B. Enzyme activity profiles in an overwintering population of freeze-tolerant larvae of the gall fly, Eurosta solidaginis . J Comp Physiol B 164, 247–255 (1994). https://doi.org/10.1007/BF00354086

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