Small membrane-associated gtp-binding proteins of catecholamine-secreting cells |
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| Institution: | 2. The National Cancer Institute, Bethesda, MDUSA;3. Lombardi Cancer Research Center, Georgetown University Medical Center, Washington, D.C., U.S.A.;1. Department of Pharmaceutics, School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi, Delhi 110062, India;2. Microbial and Pharmaceutical Biotechnology Laboratory, Jamia Hamdard, New Delhi 110062, India;3. Product Development Cell, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India;4. Division of Radiopharmaceuticals and Radiation Biology, Institute of Nuclear Medicine and Allied Sciences, Brig. SK Mazumdar Road, Delhi 110 054, India;5. Division of Biochemistry, SKUAST, Jammu 180009, India;6. Department of Biochemistry, College of Science, King Saudi University, P.O. Box. 2455, Riyadh 11451, Saudi Arabia;7. Botany and Microbiology Department, College of Science, King Saudi University, P.O. Box. 2460, Riyadh 11451, Saudi Arabia |
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| Abstract: | - 1.1. Four GTP-binding proteins (23–27 kDa) were identified in membranes from PC12 cells by α32P]GTP binding to nitrocellulose blots of SDS-polyacrylamide gels.
- 2.2. The GTP-binding proteins remained associated with membranes during stimulation of intact cells by K+-depolarization or even after addition of C2+to digitonin-permeabilized cells.
- 3.3. By two-dimensional gel electrophoresis, six GTP-binding proteins were resolved and based on their mobility, their phosphorylation state appeared independent of Ca2+.
- 4.4. Fractionation of PC12 membranes showed that these GTP-binding proteins were broadly distributed in post-nuclear membranes with the plasma membranes containing the highest specific GTP-binding activity.
- 5.5. Membrane fractions from bovine adrenal medulla contain similar GTP-binding proteins with GTP-binding intensity also being highest in the plasma membrane.
- 6.6. The GTP-binding proteins could be concentrated in the detergent-rich fraction upon Triton X-114 phase separation.
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