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Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein.
Open accessAbstract
A method is described for radioiodination to high specific activity of fixed and stained proteins within sodium dodecyl sulfate-polyacrylamide gels, without elution of the proteins from the gel. Following radioiodination, the proteins can be removed from the gel by trypsin treatment and the peptides analyzed. This procedure offers a means to structurally compare the proteins of multicomponent systems when purification of each component to homogeneity is unfeasible. Using this technique, we have compared the tryptic peptides of all the major protein components of Moloney and Rauscher leukemia viruses using only 50 to 100 microgram of total protein from each virus. Additionally, we have analyzed the membrane proteins of Dictyostelium discoideum at various stages in development. The validity of the technique and its value as a tool for comparative studies and identification of precursor-product relationships is discussed.
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Citations
Cites
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
LAEMMLI, U. K. (1970) · Nature · doi:10.1038/227680a0 · Cited by 249,961
Cited by
Kennedy, M. B., Bennett, M. K., & Erondu, N. E. (1983) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.80.23.7357 · Cited by 610
Zannis, V. I., & Breslow, J. L. (1981) · Biochemistry · doi:10.1021/bi00507a059 · Cited by 563
Bennett, M. K., Erondu, N. E., & Kennedy, M. B. (1983) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(17)44239-6 · Cited by 481
Multiple phosphorylation sites in mammalian neurofilament polypeptides.
Julien, J. P., & Mushynski, W. E. (1982) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(18)34041-9 · Cited by 379
Structural relationships between clathrin assembly proteins from the Golgi and the plasma membrane.
Ahle, S., Mann, A., Eichelsbacher, U., & Ungewickell, E. (1988) · The EMBO Journal · doi:10.1002/j.1460-2075.1988.tb02897.x · Cited by 354
Metadata
- Authors
- Elder, J. H., Pickett, R. A., Hampton, J., & Lerner, R. A.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1977-09
- Volume
- 252
- Issue
- 18
- Pages
- 6510-6515
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1515213049
- Cited by
- 774
- References
- 1
- FWCI
- 32.07
- Type
- journal article
- Language
- en
- Open access
- hybrid
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- Elder, J. H., Pickett, R. A., Hampton, J., & Lerner, R. A. (1977). Radioiodination of proteins in single polyacrylamide gel slices. Tryptic peptide analysis of all the major members of complex multicomponent systems using microgram quantities of total protein. Journal of Biological Chemistry. https://doi.org/10.1016/s0021-9258(17)39987-8
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Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
