Milk lipids, which are largely triglycerides, exist in the form of globules which are surrounded by a membrane. This membrane originates from the apical plasma membrane, which envelops globules during their release from the cell. Within the cell, cytoplasmic lipid droplets appear to be the immediate precursors of milk lipid globules. Little is known about the intracellular origin and mechanism of growth of these cytoplasmic lipid droplets. The objective of this study is to gain information on the origin and growth of the cytoplasmic lipid droplet precursors of milk lipid globules. Specifically, the hypothesis that triglyceride containing, vesicle-like structures bleb from endoplasmic reticulum and fuse with each other to form cytoplasmic lipid droplets will be tested. A further prediction of the hypothesis, that these triglyceride containing vesicles fuse with cytoplasmic lipid droplets to provide triglycerides and surface coat material necessary for growth of cytoplasmic droplets, will also be tested. Experiments to determine whether proteins of the coat material on surfaces of cytoplasmic lipid droplets may be involved in interaction of droplets with the apical plasma membrane are proposed. Approaches to these questions will involve application of ultrastructural, immunologic and biochemical techniques to tissues and to isolated cell fractions. These studies focus on one aspect of the overall goal of this research program, which is to ultimately understand what controls the rate of synthesis and secretion of the constituents of milk.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM031244-06
Application #
3279171
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1982-01-01
Project End
1988-11-30
Budget Start
1986-12-01
Budget End
1987-11-30
Support Year
6
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Virginia Polytechnic Institute and State University
Department
Type
Earth Sciences/Resources
DUNS #
003137015
City
Blacksburg
State
VA
Country
United States
Zip Code
24060
Keenan, T W; Zierdt, C H (1994) Lipid biosynthesis by axenic strains of Blastocystis hominis. Comp Biochem Physiol Biochem Mol Biol 107:525-31
Ghosal, D; Shappell, N W; Keenan, T W (1994) Endoplasmic reticulum lumenal proteins of rat mammary gland. Potential involvement in lipid droplet assembly during lactation. Biochim Biophys Acta 1200:175-81
Keon, B H; Ankrapp, D P; Keenan, T W (1994) Cytosolic lipoprotein particles from milk-secreting cells contain fatty acid synthase and interact with endoplasmic reticulum. Biochim Biophys Acta 1215:327-36
Moreau, P; Cassagne, C; Keenan, T W et al. (1993) Ceramide excluded from cell-free vesicular lipid transfer from endoplasmic reticulum to Golgi apparatus. Evidence for lipid sorting. Biochim Biophys Acta 1146:9-16
Ghosal, D; Ankrapp, D; Keenan, T W (1993) Low molecular mass GTP-binding proteins are secreted from mammary epithelial cells in association with lipid globules. Biochim Biophys Acta 1168:299-306
Keon, B H; Ghosal, D; Keenan, T W (1993) Association of cytosolic lipids with fatty acid synthase from lactating mammary gland. Int J Biochem 25:533-43
Taranto, P A; Keenan, T W; Potts, M (1993) Rehydration induces rapid onset of lipid biosynthesis in desiccated Nostoc commune (Cyanobacteria). Biochim Biophys Acta 1168:228-37
Keenan, T W; Huang, C M; Zierdt, C H (1992) Comparative analysis of lipid composition in axenic strains of Blastocystis hominis. Comp Biochem Physiol B 102:611-5
Keenan, T W; Dylewski, D P; Ghosal, D et al. (1992) Milk lipid globule precursor release from endoplasmic reticulum reconstituted in a cell-free system. Eur J Cell Biol 57:21-9
Szczepaniak, A; Huang, D; Keenan, T W et al. (1991) Electrostatic destabilization of the cytochrome b6f complex in the thylakoid membrane. EMBO J 10:2757-64

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