We propose to study biogenetic relations and functional interactions among the membranes of those intracellular compartments that together constitute the secretory pathway (or the endoplasmic reticulum-plasmalemma pathway). These compartments include the endoplasmic reticulum, a series of Golgi compartments, secretory vacuoles (or granules) and different carrier vesicles active at different junctions along this pathway which leads to discharge sites on the plasmalemma. Whenever needed, we propose to improve cell fractionation procedures by using specific ligands (e.g. antibodies or lectins) - rather than general physical properties - for the isolation of subcellular components. The cell fractions obtained will be used to isolate and purify their membranes. The proteins of these membranes will be analyzed and identified by appropriate procedures e.g., gel electrophoresis, immuneoverlays, immunoprecipitation, other biochemically specific interactions, that can be carried out on gels or gel transfers. The cell types selected for systematic investigation are the murine erythroblasts (normal or transformed) and the rat hepatocyte. The membrane proteins of primary interest are: 1) the murine glycophorins (for erythroblasts) and 2) the secretory component (for hepatocyte). Other membrane proteins will be selected in time. In each case, the pathway followed by these proteins from their sites of synthesis to their sites of final functional residence (or discharge) will be followed and the compartments involved in each postranslocational modification will be identified. With this information secured, attempts will be made to analyze the mechanisms involved in the control of intracellular vesicular traffic and - in conjunction with it - in the control of chemical specificity among interacting membranes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM027303-10
Application #
3274708
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1979-12-01
Project End
1990-03-15
Budget Start
1988-12-01
Budget End
1990-03-15
Support Year
10
Fiscal Year
1989
Total Cost
Indirect Cost
Name
Yale University
Department
Type
Schools of Medicine
DUNS #
082359691
City
New Haven
State
CT
Country
United States
Zip Code
06520
Larkin, J M; Palade, G E (1991) Transcytotic vesicular carriers for polymeric IgA receptors accumulate in rat hepatocytes after bile duct ligation. J Cell Sci 98 ( Pt 2):205-16
Sztul, E; Kaplin, A; Saucan, L et al. (1991) Protein traffic between distinct plasma membrane domains: isolation and characterization of vesicular carriers involved in transcytosis. Cell 64:81-9
Ulmer, J B; Dolci, E D; Palade, G E (1989) Glycophorin expression in murine erythroleukaemia cells. J Cell Sci 92 ( Pt 2):163-71
Dolci, E D; Palade, G E (1989) Ontogenetic expression of the murine erythrocyte glycophorins. J Cell Sci 93 ( Pt 1):191-7
Ulmer, J B; Palade, G E (1989) Anomalies in the translocation and processing of glycophorin precursors in murine erythroleukemia cells. J Biol Chem 264:1084-91
Saraste, J; Bronson, M; Palade, G E et al. (1988) Organelle-specific antibodies: production of antibodies to Golgi subcompartments. Prog Clin Biol Res 270:129-39
Saraste, J; Palade, G E; Farquhar, M G (1987) Antibodies to rat pancreas Golgi subfractions: identification of a 58-kD cis-Golgi protein. J Cell Biol 105:2021-9
Merisko, E M; Fletcher, M; Palade, G E (1986) The reorganization of the Golgi complex in anoxic pancreatic acinar cells. Pancreas 1:95-109
Saraste, J; Palade, G E; Farquhar, M G (1986) Temperature-sensitive steps in the transport of secretory proteins through the Golgi complex in exocrine pancreatic cells. Proc Natl Acad Sci U S A 83:6425-9
Larkin, J M; Sztul, E S; Palade, G E (1986) Phosphorylation of the rat hepatic polymeric IgA receptor. Proc Natl Acad Sci U S A 83:4759-63

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