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. 1978 Jun;61(6):961–966. doi: 10.1104/pp.61.6.961

Purification and Characterization of the Photoreducible b-type Cytochrome from Dictyostelium discoideum 1

Katsushi Manabe 1,2, Kenneth L Poff 1
PMCID: PMC1092021  PMID: 16660435

Abstract

The photoreducible cytochrome (Cyt) b from Dictyostelium discoideum was purified by differential precipitation with ammonium sulfate and chromatography over Sephadex G-100, diethylaminoethyl-cellulose, and calcium phosphate. The purified Cyt is composed of a single subunit of 15,000 daltons including a noncovalently bound protohaem, and exhibits in the reduced form alpha absorption bands at 555.5 and 560 nm at room temperature and 551 and 558.5 nm at 77 K. This Cyt is similar in some respects to Cyt b557.5 from complex II of beef heart mitochondria, and to Cyt b555 from the microsomal fraction of mung bean seedlings. Photoreduction by blue light of the purified Cyt b requires the addition of flavin; flavoprotein isolated from D. discoideum was the most active of four flavoproteins tested in catalyzing the photoreduction while diaphorase and l-amino-acid oxidase were inactive.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. APPLEBY C. A., MORTON R. K. Crystalline cytochrome b2 and lactic dehydrogenase of yeast. Nature. 1954 Apr 24;173(4408):749–752. doi: 10.1038/173749a0. [DOI] [PubMed] [Google Scholar]
  2. Bergman K., Burke P. V., Cerdá-Olmedo E., David C. N., Delbrück M., Foster K. W., Goodell E. W., Heisenberg M., Meissner G., Zalokar M. Phycomyces. Bacteriol Rev. 1969 Mar;33(1):99–157. doi: 10.1128/br.33.1.99-157.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brain R. D., Freeberg J. A., Weiss C. V., Briggs W. R. Blue light-induced Absorbance Changes in Membrane Fractions from Corn and Neurospora. Plant Physiol. 1977 May;59(5):948–952. doi: 10.1104/pp.59.5.948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Butler W. L. Absorption spectroscopy of biological materials. Methods Enzymol. 1972;24:3–25. doi: 10.1016/0076-6879(72)24052-6. [DOI] [PubMed] [Google Scholar]
  5. Davis K. A., Hatefi Y., Poff K. L., Butler W. L. The b-type cytochromes of bovine heart mitochondria: absorption spectra, enzymatic properties, and distribution in the electron transfer complexes. Biochim Biophys Acta. 1973 Dec 14;325(3):341–356. doi: 10.1016/0005-2728(73)90196-5. [DOI] [PubMed] [Google Scholar]
  6. Delbrück M., Katzir A., Presti D. Responses of Phycomyces indicating optical excitation of the lowest triplet state of riboflavin. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1969–1973. doi: 10.1073/pnas.73.6.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  8. Jesaitis A. J., Heners P. R., Hertel R. Characterization of a Membrane Fraction Containing a b-type Cytochrome. Plant Physiol. 1977 May;59(5):941–947. doi: 10.1104/pp.59.5.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jesaitis A. J. Linear dichroism and orientation of the Phycomyces photopigment. J Gen Physiol. 1974 Jan;63(1):1–21. doi: 10.1085/jgp.63.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Muñoz V., Brody S., Butler W. L. Photoreceptor pigment for blue light responses in Neurospora crassa. Biochem Biophys Res Commun. 1974 May 7;58(1):322–327. doi: 10.1016/0006-291x(74)90930-9. [DOI] [PubMed] [Google Scholar]
  12. Muñoz V., Butler W. L. Photoreceptor Pigment for Blue Light in Neurospora crassa. Plant Physiol. 1975 Feb;55(2):421–426. doi: 10.1104/pp.55.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Poff K. L., Butler W. L. Absorbance changes induced by blue light in Phycomyces blakesleeanus and Dictyostelium discoideum. Nature. 1974 Apr 26;248(5451):799–801. doi: 10.1038/248799a0. [DOI] [PubMed] [Google Scholar]
  14. Poff K. L., Butler W. L. Spectral Characterization of the Photoreducible b-Type Cytochrome of Dictyostelium discoideum. Plant Physiol. 1975 Feb;55(2):427–429. doi: 10.1104/pp.55.2.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SHICHI H., HACKETT D. P., FUNATSU G. Studies on the b-type cytochromes from mung bean seedlings. III. The structure of cytochrome b-555. J Biol Chem. 1963 Mar;238:1156–1161. [PubMed] [Google Scholar]
  16. SHICHI H., HACKETT D. P. Studies on the b-type cytochromes from mung bean seedlings. I. Purification of cytochromes b-555 and b-561. J Biol Chem. 1962 Sep;237:2955–2958. [PubMed] [Google Scholar]
  17. SHICHI H., HACKETT D. P. Studies on the b-type cytochromes from mung bean seedlings. II. Some properties of cytochromes b-555 and b-561. J Biol Chem. 1962 Sep;237:2959–2964. [PubMed] [Google Scholar]
  18. SIEGELMAN H. W., FIRER E. M. PURIFICATION OF PHYTOCHROME FROM OAT SEEDLINGS. Biochemistry. 1964 Mar;3:418–423. doi: 10.1021/bi00891a019. [DOI] [PubMed] [Google Scholar]
  19. Schmidt W., Butler W. L. Flavin-mediated photoreactions in artificial systems: a possible model for the blue-light photoreceptor pigment in living systems. Photochem Photobiol. 1976 Jul;24(1):71–75. doi: 10.1111/j.1751-1097.1976.tb06799.x. [DOI] [PubMed] [Google Scholar]
  20. Schmidt W., Butler W. L. Light-induced absorbance changes in cell-free extracts of Neurospora crassa. Photochem Photobiol. 1976 Jul;24(1):77–80. doi: 10.1111/j.1751-1097.1976.tb06800.x. [DOI] [PubMed] [Google Scholar]
  21. Schmidt W., Hart J., Filner P., Poff K. L. Specific inhibition of phototropism in corn seedlings. Plant Physiol. 1977 Nov;60(5):736–738. doi: 10.1104/pp.60.5.736. [DOI] [PMC free article] [PubMed] [Google Scholar]

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