Human milk bile salt-activated lipase (BAL) is a physiologically important enzyme in the intestinal fat digestion processes of infants. The goal of this project is to understand the structure and function relationships of this enzyme. The aspects of the proposed studies are summarized in the following. (1) We propose to determine the sequence of its cDNA. This will provide a structural framework for the functional interpretations. (2) The identification and structure determination of the active site will help define the nature and location of functional groups essential for the functional interpretations. (3) The further understanding of the enzyme kinetics will be instrumental for gaining an understanding of the catalytic mechanism of BAL. (4) The cloned human milk BAL cDNA will be expressed in mammalian cell culture which will be used to carry out site-directed mutagenesis experiments. This approach will permit the pin-point examination of the roles of functionally important residues in BAL by muta- genesis experiments. (5) The structure-function information generated for human milk BAL will also facilitate a comparative study on the pancreatic BAL. These studies may include the determination of cDNA structure and the kinetic studies of pancreatic BAl.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD023472-02
Application #
3323665
Study Section
Physiological Chemistry Study Section (PC)
Project Start
1989-04-01
Project End
1994-03-31
Budget Start
1990-04-01
Budget End
1991-03-31
Support Year
2
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Oklahoma Medical Research Foundation
Department
Type
DUNS #
937727907
City
Oklahoma City
State
OK
Country
United States
Zip Code
73104
Wang, C S; Downs, D; Dashti, A et al. (1996) Isolation and characterization of recombinant human apolipoprotein C-II expressed in Escherichia coli. Biochim Biophys Acta 1302:224-30
Wang, C S; Dashti, A; Jackson, K W et al. (1995) Isolation and characterization of human milk bile salt-activated lipase C-tail fragment. Biochemistry 34:10639-44
Wang, C S (1994) Probing of active site structure of lipoprotein lipase: contribution of activation entropy in the catalysis. Biochim Biophys Acta 1212:67-72
Downs, D; Xu, Y Y; Tang, J et al. (1994) Proline-rich domain and glycosylation are not essential for the enzymic activity of bile salt-activated lipase. Kinetic studies of T-BAL, a truncated form of the enzyme, expressed in Escherichia coli. Biochemistry 33:7979-85
Wang, C S; Bass, H; Whitmer, R et al. (1993) Effects of albumin and apolipoprotein C-II on the acyl-chain specificity of lipoprotein lipase catalysis. J Lipid Res 34:2091-8
Wang, C S; Hartsuck, J A (1993) Bile salt-activated lipase. A multiple function lipolytic enzyme. Biochim Biophys Acta 1166:1-19
Wang, C S; Hartsuck, J; McConathy, W J (1992) Structure and functional properties of lipoprotein lipase. Biochim Biophys Acta 1123:1-17
McConathy, W J; Gesquiere, J C; Bass, H et al. (1992) Inhibition of lipoprotein lipase activity by synthetic peptides of apolipoprotein C-III. J Lipid Res 33:995-1003
Wang, C S (1991) Structure and functional properties of apolipoprotein C-II. Prog Lipid Res 30:253-8
Baba, T; Downs, D; Jackson, K W et al. (1991) Structure of human milk bile salt activated lipase. Biochemistry 30:500-10

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