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Purification of the regulatory component of adenylate cyclase.
Open accessAbstract
The regulatory component (G/F) of adenylate cyclase [ATP pyrophosphate-lyase (cyclizing), EC 4.6.1.1] from rabbit liver plasma membranes has been purified essentially to homogeneity. The purification was accomplished by three chromatographic procedures in sodium cholate-containing solutions, followed by three steps in Lubrol-containing solutions. The specific activity of G/F was enriched 2000-fold from extracts of membranes to 3-4 mumol x min-1 x mg-1 (reconstituted adenylate cyclase activity). Purified G/F reconstitutes guanine nucleotide-, fluoride-, and hormone-stimulated adenylate cyclase activity in the adenylate cyclase-deficient variant of S49 murine lymphoma cells. G/F also recouples hormonal stimulation of the enzyme in the uncoupled variant of S49. Preparations of pure G/F contain three polypeptides with approximate molecular weights of 52,000, 45,000, and 35,000. The active G/F protein behaves as a multisubunit complex of these polypeptides. Treatment of G/F with [32P]NAD+ and cholera toxin covalently labels the molecular weight 52,000 and 45,000 polypeptides with 32P.
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Cited by
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Taparowsky, E., Shimizu, K., Goldfarb, M., & Wigler, M. (1983) · Cell · doi:10.1016/0092-8674(83)90390-2 · Cited by 527
Weinstein, L. S., Yu, S., Warner, D. R., & Liu, J. (2001) · Endocrine Reviews · doi:10.1210/edrv.22.5.0439 · Cited by 420
Fluoroaluminates activate transducin‐GDP by mimicking the γ‐phosphate of GTP in its binding site
Bigay, J., Deterre, P., Pfister, C., & Chabre, M. (1985) · FEBS Letters · doi:10.1016/0014-5793(85)80004-1 · Cited by 417
Bacterial toxins: cellular mechanisms of action
Middlebrook, J. L., & Dorland, R. B. (1984) · Microbiological Reviews · doi:10.1128/mmbr.48.3.199-221.1984 · Cited by 357
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Barrowman, M. M., Cockcroft, S., & Gomperts, B. D. (1986) · Nature · doi:10.1038/319504a0 · Cited by 337
Metadata
- Authors
- Northup, J. K., Sternweis, P. C., Smigel, M. D., Schleifer, L. S., Ross, E. M., & Gilman, A. G.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1980-11
- Volume
- 77
- Issue
- 11
- Pages
- 6516-6520
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W1984080990
- Cited by
- 506
- References
- 0
- FWCI
- 17.95
- Type
- journal article
- Language
- en
- Open access
- green
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- Northup, J. K., Sternweis, P. C., Smigel, M. D., Schleifer, L. S., Ross, E. M., & Gilman, A. G. (1980). Purification of the regulatory component of adenylate cyclase. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.77.11.6516
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