Abstract

The project encompasses studies of three systems of distinct biochemical significance: lactose synthase, human serum transferrin and its cellular receptors and the enzymatic N-glycosylation of proteins. A connecting theme in the investigation is the identification of the molecular locations of the interaction sites on alpha-lactalbumin and galactosyltransferase, the two protein components of lactose synthase, and of the receptor interaction site of transferrin. Approaches will include differential kinetic labelling, covalent crosslinking with bifunctional reagents as well as the examination of fragments for inhibitory effects on the interactions. The relationship of structure and function in the individual proteins will also be investigated. Studies of protein N-glycosylation will focus on the use of peptide analogues containing modified glycosylation signal sequence as inhibitors and active site covalent labels.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM021363-12
Application #
3270436
Study Section
Biochemistry Study Section (BIO)
Project Start
1977-06-01
Project End
1990-05-31
Budget Start
1985-09-17
Budget End
1986-05-31
Support Year
12
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
University of Miami School of Medicine
Department
Type
Schools of Medicine
DUNS #
City
Miami
State
FL
Country
United States
Zip Code
33101

  Publications

Greene, Lesley H; Hamada, Daizo; Eyles, Stephen J et al. (2003) Conserved signature proposed for folding in the lipocalin superfamily. FEBS Lett 553:39-44
Greene, L H; Chrysina, E D; Irons, L I et al. (2001) Role of conserved residues in structure and stability: tryptophans of human serum retinol-binding protein, a model for the lipocalin superfamily. Protein Sci 10:2301-16
Greene, L H; Grobler, J A; Malinovskii, V A et al. (1999) Stability, activity and flexibility in alpha-lactalbumin. Protein Eng 12:581-7
Zhang, Y; Malinovskii, V A; Fiedler, T J et al. (1999) Role of a conserved acidic cluster in bovine beta1,4 galactosyltransferase-1 probed by mutagenesis of a bacterially expressed recombinant enzyme. Glycobiology 9:815-22
Chandra, N; Brew, K; Acharya, K R (1998) Structural evidence for the presence of a secondary calcium binding site in human alpha-lactalbumin. Biochemistry 37:4767-72
Pike, A C; Brew, K; Acharya, K R (1996) Crystal structures of guinea-pig, goat and bovine alpha-lactalbumin highlight the enhanced conformational flexibility of regions that are significant for its action in lactose synthase. Structure 4:691-703
Malinovskii, V A; Tian, J; Grobler, J A et al. (1996) Functional site in alpha-lactalbumin encompasses a region corresponding to a subsite in lysozyme and parts of two adjacent flexible substructures. Biochemistry 35:9710-5
Grobler, J A; Wang, M; Pike, A C et al. (1994) Study by mutagenesis of the roles of two aromatic clusters of alpha-lactalbumin in aspects of its action in the lactose synthase system. J Biol Chem 269:5106-14
Grobler, J A; Rao, K R; Pervaiz, S et al. (1994) Sequences of two highly divergent canine type c lysozymes: implications for the evolutionary origins of the lysozyme/alpha-lactalbumin superfamily. Arch Biochem Biophys 313:360-6
Brandt, N R; Caswell, A H; Brandt, T et al. (1992) Mapping of the calpain proteolysis products of the junctional foot protein of the skeletal muscle triad junction. J Membr Biol 127:35-47

Showing the most recent 10 out of 31 publications