We have investigated the interaction of lipoproteins with liposomes to form recombinant particles. A number of lipoprotein fractions (VLDL, IDL, LDL, and HDL) all disrupt liposome structure by an essentially irreversible and quasistoichiometric process. In the case of HDL, the major apoprotein, A-I, recombines with dimyristoyl phosphatidyl choline vesicles 40:1 lipid-protein to form discs approximately 100 angstrom unit in diameter and 32 angstrom unit in thickness, with protein on the rim. These structural results were obtained by a combination of neutron scattering, electron microscopy, column chromatography, and fluorescence techniques. With dipalmitoyl phosphatidylcholine, A-I also forms what we term """"""""vesicular recombinant"""""""" particles in a process which may relate to physiological mechanisms by which proteins are assembled into membranes and lipoproteins. To study this process we have developed a technique called """"""""phase transition release"""""""" (PTR) which is also being applied to study incorporation of tubulin into membranes. Lipoproteins were labelled with the fluorescent lipid 3,3 dioctadecylindo-carbocyanine for studies of interaction will cell surface lipoprotein receptors. The lipoproteins are also being labelled with NBD lipids for two-color fluorescence identification of cells in atheroscleroic plaques. Statistical and more general mechanical algorithms (HAL, HALP, HALCO) were devised for evaluating amphipathic helical structures and more general structure-function relationships in proteins and peptides. This is being used to define issues of structure and immunogenicity with respect to HLA antigens and to the envelope polyprotein of HIV. Lipid membrane systems and human cell isolates are being used experimentally to investigate the interaction between characterized synthetic antigenic peptides and T-cells in the recognition process. The results may have application to the design of vaccines.

Agency
National Institute of Health (NIH)
Institute
Division of Cancer Biology And Diagnosis (NCI)
Type
Intramural Research (Z01)
Project #
1Z01CB008341-10
Application #
3939287
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
10
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Cancer Biology and Diagnosis
Department
Type
DUNS #
City
State
Country
United States
Zip Code