Kinetics of a novel cytosolic protein during the onset of renal epithelial cell growth
| Autor: | Sreedharan Kartha, F. G. Toback, M. M. Walsh-Reitz, W. A. Franklin, Stephen L. Gluck, H. N. Aithal, K. M. Knigge |
|---|---|
| Rok vydání: | 1988 |
| Předmět: |
Physiology
medicine.medical_treatment Blotting Western Immunocytochemistry Enzyme-Linked Immunosorbent Assay Biology Kidney Monospecific antibody Epithelium Cell Line Cytosol Chlorocebus aethiops medicine Animals Chromatography High Pressure Liquid Molecular mass Cell growth Growth factor Glyceraldehyde-3-Phosphate Dehydrogenases Proteins Epithelial Cells Immunohistochemistry Molecular biology Enzyme assay Molecular Weight Kinetics Biochemistry Cell culture Chromatography Gel Potassium biology.protein Cell Division |
| Zdroj: | American Journal of Physiology-Renal Physiology. 255:F868-F873 |
| ISSN: | 1522-1466 1931-857X |
| DOI: | 10.1152/ajprenal.1988.255.5.f868 |
| Popis: | Exposure of monkey kidney epithelial cells (BSC-1 line) to medium with a reduced K concentration (3.2 mM) stimulated growth and transiently activated glyceraldehyde-3-phosphate dehydrogenase (G3PD). The increase in enzyme activity was mediated by a cytosolic modifier protein that was purified using affinity and size-exclusion chromatography, and anion-exchange high-performance liquid chromatography. The apparent molecular mass of the protein was 62 kDa. A monospecific antibody to the protein was prepared from rabbit antiserum and used as an immunoprobe. Immunocytochemical staining and Western blotting revealed that the protein was a normal constituent of the cytosol and that it accumulated in cells exposed to low-K medium. A quantitative enzyme-linked immunosorbent assay showed that the amount of modifier protein increased progressively for up to 2 h in cells exposed to low-K medium, and then returned to the control value, a kinetic profile similar to that observed for G3PD activity. These results indicate that the modifier protein is a constituent of renal epithelial cells and accumulates transiently in the cytosol where it could regulate G3PD activity during the onset of growth induced by the low-K mitogenic signal. |
| Databáze: | OpenAIRE |
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