Radiation inactivation analysis of rat liver microsomal glucose-6-phosphatase.
Radiation inactivation analysis was utilized to estimate the sizes of the units catalyzing the various activities of hepatic microsomal glucose-6-phosphatase. This technique revealed that the target molecular weights for mannose-6-P phosphohydrolase, glucose-6-P phosphohydrolase, and carbamyl-P:glucose phosphotransferase activities were all about Mr 75,000. These results are consistent with the widely held view that all of these activities are catalyzed by the same protein or proteins. Certain observations indicate that the molecular organization of microsomal glucose-6-phosphatase is better described by the conformational hypothesis which envisions the enzyme as a single covalent structure rather than by the substrate transport model which requires the participation of several physically separate polypeptides. These include the findings: 1) that the target sizes for glucose-6-P phosphohydrolase and carbamyl-P:glucose phosphotransferase activities were not larger than that for mannose-6-P phosphohydrolase in intact microsomes and 2) that the target size for glucose-6-P phosphohydrolase in disrupted microsomes was not less than that observed in intact microsomes. These findings are most consistent with a model for glucose-6-phosphatase of a single polypeptide or a disulfide-linked dimer which spans the endoplasmic reticulum with the various activities of this multifunctional enzyme residing in distinct protein domains.
Ness
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GC
Department of Biochemistry and Molecular Biology, College of Medicine, University of South Florida, Tampa 33612.
Sukalski
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KA
Sample
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Pendleton
L C
LC
McCreery
M J
MJ
Nordlie
R C
RC
D000818:Animals; D003840:Deoxycholic Acid / Q000494:pharmacology; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology; D004307:Dose-Response Relationship, Radiation; D005952:Glucose-6-Phosphatase / Q000528:radiation effects*; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D011487:Protein Conformation; D051381:Rats; D011919:Rats, Inbred Strains; D013329:Structure-Activity Relationship
D003840:Deoxycholic Acid; D005952:Glucose-6-Phosphatase