Leucine-Specific Binding of Photoreactive Leu7-MAP to a High Molecular Weight Protein on the Plasma Membrane of the Isolated Rat Hepatocyte
| Autor: | John J. Wert, Giovanni Miotto, G E Mortimore, M Kadowaki, Rina Venerando |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
Male
Proteolysis Molecular Sequence Data Biophysics Peptide Biochemistry Dithiothreitol Iodine Radioisotopes chemistry.chemical_compound Leucine AMINO-ACIDS HEPATIC PROTEOLYSIS DEGRADATION DEPRIVATION Liver medicine Animals AMINO-ACIDS Amino Acid Sequence DEPRIVATION Molecular Biology Cells Cultured chemistry.chemical_classification medicine.diagnostic_test Cell Membrane Membrane Proteins Biological activity Cell Biology DEGRADATION Rats Molecular Weight Kinetics Cytosol Membrane medicine.anatomical_structure Liver chemistry Rats Inbred Lew Hepatocyte Intercellular Signaling Peptides and Proteins Electrophoresis Polyacrylamide Gel HEPATIC PROTEOLYSIS Peptides |
| Zdroj: | Biochemical and Biophysical Research Communications. 203:200-208 |
| ISSN: | 0006-291X |
| DOI: | 10.1006/bbrc.1994.2168 |
| Popis: | Leu8-MAP (Multiple Antigen Peptide) is an effective inhibitor of macroautophagy and proteolysis in the isolated rat hepatocyte, having an apparent Km (0.1 mM) equaling leucine. Since it is not transported into the cytosolic compartment, it very likely mediates its effect through a plasma membrane site. In an attempt to identify the site we photoreacted intact cells with a biologically active, iodinatable azide derivative of Leu7-MAP. A approximately 340,000 M(r) protein whose labeling was protected 83% with 20 mM Leu was found in plasma membrane fractions when electrophoresed in 7.5-20% gradient gels under nonreducing conditions; addition of 20 mM dithiothreitol generated smaller m.w. products, possibly subunits, of consistent size. No specific labeling was observed with photoreactive derivatives of Ile7-MAP or Val7-MAP. |
| Databáze: | OpenAIRE |
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