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 Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK026564-07
Application #
3227956
Study Section
Biochemistry Study Section (BIO)
Project Start
1980-04-01
Project End
1990-03-31
Budget Start
1986-04-01
Budget End
1987-03-31
Support Year
7
Fiscal Year
1986
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

Marcus, F; Harrsch, P B (1990) Amino acid sequence of spinach chloroplast fructose-1,6-bisphosphatase. Arch Biochem Biophys 279:151-7
Ladror, U S; Latshaw, S P; Marcus, F (1990) Spinach cytosolic fructose-1,6-bisphosphatase. Purification, enzyme properties and structural comparisons. Eur J Biochem 189:89-94
Ladror, U S (1990) Calculating percent identity between protein or DNA sequences with a word processor. Protein Seq Data Anal 3:267-71
Valaitis, A P; Foe, L G; Kwiatkowska, D et al. (1989) The sites of phosphorylation of rabbit brain phosphofructo-1-kinase by cyclic AMP-dependent protein kinase. Biochim Biophys Acta 995:187-94
Marcus, F; Rittenhouse, J; Moberly, L et al. (1988) Yeast (Saccharomyces cerevisiae) fructose-1,6-bisphosphatase. Properties of phospho and dephospho forms and of two mutants in which serine 11 has been changed by site-directed mutagenesis. J Biol Chem 263:6058-62
Banas, T; Gontero, B; Drews, V L et al. (1988) Reactivity of the thiol groups of Escherichia coli phosphofructo-1-kinase. Biochim Biophys Acta 957:178-84
Marcus, F; Fickenscher, K (1988) Amino-terminal sequence of spinach chloroplast fructose-1,6-bisphosphatase. Arch Biol Med Exp (Santiago) 21:117-21
Marcus, F; Moberly, L; Latshaw, S P (1988) Comparative amino acid sequence of fructose-1,6-bisphosphatases: identification of a region unique to the light-regulated chloroplast enzyme. Proc Natl Acad Sci U S A 85:5379-83
Yuan, X H; Kwiatkowska, D; Kemp, R G (1988) Inorganic pyrophosphate: fructose-6-phosphate 1-phosphotransferase of the potato tuber is related to the major ATP-dependent phosphofructokinase of E. coli. Biochem Biophys Res Commun 154:113-7
Valaitis, A P; Kwiatkowska, D; Krishnaraj, R et al. (1988) Identification of the AMP binding sites of rabbit phosphofructo-1-kinase isozymes B and C. Biochim Biophys Acta 956:232-42

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