This grant will support research in two areas: lipoprotein structure and molecular immunology. The common thread is our interest in the structure of functional macromolecular complexes formed between proteins and lipids and receptors, and between antibodies, antigen, and complement. The goals of the lipoprotein projects are to (1) determine the conformation of apolipoprotein B on the surface of the low density lipoprotein, and the location of the site on apo B recognized by the receptor; (2) to determine the number of apo B48 molecules on the chylomicrons, and to examine the confomation of apo B100 on the VLDL; (3) to characterize the structure of recombinant apolipoprotein(a) and its complex with LDL which forms the highly atherogenic Lp(a). The goals of the molecular immunology projects are (1) to determine the nature of the conformational changes through which IgM acquires the ability to bind and activate complement; (2) to develop a new technique we call 'targeted immunogenesis' to form monoclonal antibodies to selected regions on a protein. These monoclonals will be used as probes of IgM structure and function. (3) to study protein protein interactions by equilibrium dialysis, employing a special membrane not previously used for such studies. This technique will then be employed in both the lipoprotein and immunoglobulin projects. The methodology includes analytical ultracentrifugation, circular dichroism, fluorescence, and electron microscopy, as well as the production of monoclonal antibodies and recombinant DNA techniques. These projects are related to problems of health; thus (1) high concentrations of chylomicron and VLDL remnants remaining after lipolysis, and especially high concentrations of LDL, are closely associated with the development of the atherosclerotic lesion, and associated heart attack, senility and stroke; (2) the triggering of complement and cells by immune complexes and antigens is a critical feature of the immune defense against bacterial and viral challenge.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM013914-27
Application #
3268555
Study Section
Biophysical Chemistry Study Section (BBCB)
Project Start
1976-09-01
Project End
1995-08-31
Budget Start
1991-09-01
Budget End
1992-08-31
Support Year
27
Fiscal Year
1991
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Type
Schools of Arts and Sciences
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Schumaker, V N; Phillips, M L; Chatterton, J E (1994) Apolipoprotein B and low-density lipoprotein structure: implications for biosynthesis of triglyceride-rich lipoproteins. Adv Protein Chem 45:205-48
Phillips, M L; Tao, M H; Morrison, S L et al. (1994) Human/mouse chimeric monoclonal antibodies with human IgG1, IgG2, IgG3 and IgG4 constant domains: electron microscopic and hydrodynamic characterization. Mol Immunol 31:1201-10
Phillips, M L; Lembertas, A V; Schumaker, V N et al. (1993) Physical properties of recombinant apolipoprotein(a) and its association with LDL to form an LP(a)-like complex. Biochemistry 32:3722-8
Lane, P D; Schumaker, V N; Tseng, Y et al. (1991) Isolation of human complement subcomponents C1r and C1s in their unactivated, proenzyme forms. J Immunol Methods 141:219-26
Schumaker, V N; Phillips, M L; Hanson, D C (1991) Dynamic aspects of antibody structure. Mol Immunol 28:1347-60
Poon, P H; Schumaker, V N (1991) Measurement of macromolecular interactions between complement subcomponents C1q, C1r, C1s, and immunoglobulin IgM by sedimentation analysis using the analytical ultracentrifuge. J Biol Chem 266:5723-7
Chatterton, J E; Phillips, M L; Curtiss, L K et al. (1991) Mapping apolipoprotein B on the low density lipoprotein surface by immunoelectron microscopy. J Biol Chem 266:5955-62
Phillips, M L; Oi, V T; Schumaker, V N (1990) Electron microscopic study of ring-shaped, bivalent hapten, bivalent antidansyl monoclonal antibody complexes with identical variable domains but IgG1, IgG2a and IgG2b constant domains. Mol Immunol 27:181-90
Phillips, M L; Schumaker, V N (1989) Conformation of apolipoprotein B after lipid extraction of low density lipoproteins attached to an electron microscope grid. J Lipid Res 30:415-22
Schumaker, V N; Tseng, Y; Poon, P H et al. (1989) Spontaneous activation of reconstituted and serum C1 and the role of C1-inhibitor. Behring Inst Mitt :102-10

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