Abstract

The project continues our structural studies of the two key regulatory enzymes of carbohydrate metabolism, phosphofructokinase (PFK) and fructose 1,6-bisphosphatase (FbPase). The research, which involves a major collaborative effort of three laboratories, includes the following: (1) The primary sequence of rabbit muscle PFK, which is almost complete, will be finished and predictions concerning three dimensional organization will be tested by cross-linking experiments. (2) Sequences around functional residues of muscle PFK will be determined by covalent modification techniques. (3) Predictions concerning the evolution of allosteric sites of PFK will be evaluated by functional site identification and partial sequence analysis of E. coli and yeast (Saccharomyces cerevisiae) PFK. (4) Functional site identification and sequence analysis of E. coli FbPase, and an analysis of the phosphorylation site(s) of Saccharomyces cerevisiae FbPase will be compared to data obtained with mammalian FbPases. (5) The sequence of skeletal muscle FbPase, an enzyme with increased sensitivity to AMP inhibition, will be determined and compared to the kidney and liver enzyme. These studies provide structural correlates to our knowledge of allosteric interactions and is of interest to our overall understanding of enzyme regulatory mechanisms.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases (NIADDK)
Type
Research Project (R01)
Project #
2R01AM026564-06
Application #
3151646
Study Section
Biochemistry Study Section (BIO)
Project Start
1980-04-01
Project End
1990-03-31
Budget Start
1985-04-01
Budget End
1986-03-31
Support Year
6
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Rosalind Franklin University of Medicine & Sci
Department
Type
Schools of Medicine
DUNS #
069501252
City
North Chicago
State
IL
Country
United States
Zip Code
60064

  Publications

Kemp, R G; Fox, R W; Latshaw, S P (1987) Amino acid sequence at the citrate allosteric site of rabbit muscle phosphofructokinase. Biochemistry 26:3443-6
Valaitis, A P; Foe, L G; Kemp, R G (1987) Desensitization of muscle phosphofructokinase to ATP inhibition by removal of a carboxyl-terminal heptadecapeptide. J Biol Chem 262:5044-8
Bezares, G; Eyzaguirre, J; Hinrichs, M V et al. (1987) Isolation and sequence determination of an active site peptide of rabbit muscle pyruvate kinase. Arch Biochem Biophys 253:133-7
Fickenscher, K; Scheibe, R; Marcus, F (1987) Amino acid sequence similarity between malate dehydrogenases (NAD) and pea chloroplast malate dehydrogenase (NADP). Eur J Biochem 168:653-8
Latshaw, S P; Bazaes, S; Randolph, A et al. (1987) Identification of highly reactive cysteinyl and methionyl residues of rabbit muscle phosphofructokinase. J Biol Chem 262:10672-7
Rittenhouse, J; Harrsch, P B; Kim, J N et al. (1986) Amino acid sequence of the phosphorylation site of yeast (Saccharomyces cerevisiae) fructose-1,6-bisphosphatase. J Biol Chem 261:3939-43
Marcus, F; Ureta, T (1986) Amino acid sequence homology between yeast hexokinases and rat hexokinase C. Biochem Biophys Res Commun 139:714-9
Marcus, F; Gontero, B; Harrsch, P B et al. (1986) Amino acid sequence homology among fructose-1,6-bisphosphatases. Biochem Biophys Res Commun 135:374-81
Marcus, F (1985) Preferential cleavage at aspartyl-prolyl peptide bonds in dilute acid. Int J Pept Protein Res 25:542-6
Harrsch, P B; Kim, Y; Fox, J L et al. (1985) Amino acid sequence similarity between spinach chloroplast and mammalian gluconeogenic fructose-1,6-bisphosphatase. Biochem Biophys Res Commun 133:520-6

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