Abstract
We have developed a phytochrome immunoaffinity purification procedure that yields undegraded oat (Avena sativa L., cv. Garry) phytochrome of greater than 98% purity within 2 hours when starting with a brushite-purified preparation. Immunoaffinity-purified phytochrome, except for its greater purity, is indistinguishable from conventionally purified phytochrome by gel exclusion chromatography, isoelectric focusing, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We have also used the immunoaffinity technique to purify phytochrome from crude oat extracts, and from brushite-purified pea (Pisum sativum L., cv. Alaska) and rye (Secale cereale L., cv. Balbo) preparations.
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Selected References
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- Correll D. L., Edwards J. L. The aggregation States of phytochrome from etiolated rye and oat seedings. Plant Physiol. 1970 Jan;45(1):81–85. doi: 10.1104/pp.45.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cundiff S. C., Pratt L. H. Immunological and physical characterization of the products of phytochrome proteolysis. Plant Physiol. 1975 Feb;55(2):212–217. doi: 10.1104/pp.55.2.212. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gardner G., Pike C. S., Rice H. V., Briggs W. R. "Disaggregation" of phytochrome in vitro-a consequence of proteolysis. Plant Physiol. 1971 Dec;48(6):686–693. doi: 10.1104/pp.48.6.686. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hunt R. E., Pratt L. H. Phytochrome radioimmunoassay. Plant Physiol. 1979 Aug;64(2):327–331. doi: 10.1104/pp.64.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lin P. P., Varner J. E. Cyclic nucleotide phosphodiesterase in pea seedlings. Biochim Biophys Acta. 1972 Aug 28;276(2):454–474. doi: 10.1016/0005-2744(72)91007-8. [DOI] [PubMed] [Google Scholar]
- Otavsky W. I., Drysdale J. W. Recent staining artifacts with LKB "Ampholines" on gel isoelectric-focusing. Anal Biochem. 1975 May 12;65(1-2):533–536. doi: 10.1016/0003-2697(75)90538-2. [DOI] [PubMed] [Google Scholar]
- Pratt L. H., Coleman R. A. Immunocytochemical localization of phytochrome. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2431–2435. doi: 10.1073/pnas.68.10.2431. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pratt L. H. Comparative immunochemistry of phytochrome. Plant Physiol. 1973 Jan;51(1):203–209. doi: 10.1104/pp.51.1.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pratt L. H., Cundiff S. C. Spectral characterization of high-molecular-weight phytochrome. Photochem Photobiol. 1975 Feb;21(2):91–97. doi: 10.1111/j.1751-1097.1975.tb06634.x. [DOI] [PubMed] [Google Scholar]
- Pratt L. H., Kidd G. H., Coleman R. A. An immunochemical characterization of the phytochrome destruction reaction. Biochim Biophys Acta. 1974 Sep 13;365(1):93–107. doi: 10.1016/0005-2795(74)90253-0. [DOI] [PubMed] [Google Scholar]
- Rice H. V., Briggs W. R., Jackson-White C. J. Purification of oat and rye phytochrome. Plant Physiol. 1973 May;51(5):917–926. doi: 10.1104/pp.51.5.917. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Righetti P., Drysdale J. W. Isoelectric focusing in polyacrylamide gels. Biochim Biophys Acta. 1971 Apr 27;236(1):17–28. doi: 10.1016/0005-2795(71)90144-9. [DOI] [PubMed] [Google Scholar]
- Tobin E. M., Briggs W. R. Studies on the protein comformation of phytochrome. Photochem Photobiol. 1973 Dec;18(6):487–495. doi: 10.1111/j.1751-1097.1973.tb06454.x. [DOI] [PubMed] [Google Scholar]
- Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]