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. 1980 Sep;66(3):428–432. doi: 10.1104/pp.66.3.428

Fractionation of Thylakoid Membranes with the Nonionic Detergent Octyl-β-d-glucopyranoside

RESOLUTION OF CHLOROPHYLL-PROTEIN COMPLEX II INTO TWO CHLOROPHYLL-PROTEIN COMPLEXES

Edith L Camm 1, Beverley R Green 1
PMCID: PMC440647  PMID: 16661449

Abstract

The detergent octyl-β-d-glucopyranoside (30 millimolar in 2 millimolar Tris-maleate, pH 7.0) preferentially extracts complexes containing protein and chlorophylls a plus b (CP) from spinach, leaving a residue highly enriched in CP I (P700-chlorophyll a protein). Use of the detergent results in a relatively gentle extraction since little free chlorophyll is formed and since sodium dodecyl sulfate-gel electrophoresis (on 10% acrylamide) of the extract also reveals the presence of two minor chlorophyll a complexes (apparent molecular weight, 47,000 and 43,000) instead of the usual single complex. The major complex preserved is CP 64, a chlorophyll a/b complex (apparent molecular weight, 64,000) which is an oligomer of another chlorophyll a/b complex, CP 27, the light-harvesting complex (apparent molecular weight, 27,000). Dissociation of each complex reveals two polypeptides (molecular weight, 32,000 and 28,000) and limited proteolysis confirms that those of CP 64 have the same structure as those of CP 27. An additional chlorophyll a/b complex (apparent molecular weight, 29,000) is clearly separable from CP 27, and differs from it and CP 64 in having a higher chlorophyll a/b ratio and a single polypeptide (molecular weight, 29,000) which differs structurally from those of the other complexes.

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Selected References

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

  1. Apel K. The light-harvesting chlorophylla a/b.protein complex of the green alga Acetabularia mediterranea. Isolation and characterization of two subunits. Biochim Biophys Acta. 1977 Nov 17;462(2):390–402. doi: 10.1016/0005-2728(77)90137-2. [DOI] [PubMed] [Google Scholar]
  2. Bar-Nun S., Schantz R., Ohad I. Appearance and composition of chlorophyll-protein complexes I and II during chloroplast membrane biogenesis in Chlamydomonas reinhardi y-1. Biochim Biophys Acta. 1977 Mar 11;459(3):451–467. doi: 10.1016/0005-2728(77)90045-7. [DOI] [PubMed] [Google Scholar]
  3. Baron C., Thompson T. E. Solubilization of bacterial membrane proteins using alkyl glucosides and dioctanoyl phosphatidylcholine. Biochim Biophys Acta. 1975 Mar 25;382(3):276–285. doi: 10.1016/0005-2736(75)90270-9. [DOI] [PubMed] [Google Scholar]
  4. Burke J. J., Ditto C. L., Arntzen C. J. Involvement of the light-harvesting complex in cation regulation of excitation energy distribution in chloroplasts. Arch Biochem Biophys. 1978 Apr 15;187(1):252–263. doi: 10.1016/0003-9861(78)90031-0. [DOI] [PubMed] [Google Scholar]
  5. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  6. Delepelaire P., Chua N. H. Lithium dodecyl sulfate/polyacrylamide gel electrophoresis of thylakoid membranes at 4 degrees C: Characterizations of two additional chlorophyll a-protein complexes. Proc Natl Acad Sci U S A. 1979 Jan;76(1):111–115. doi: 10.1073/pnas.76.1.111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dunkley P. R., Anderson J. M. The light-harvesting chlorophyll a/b-protein complex from barley thylakoid membranes. Polypeptide composition and characterization of an oligomer. Biochim Biophys Acta. 1979 Jan 11;545(1):174–187. [PubMed] [Google Scholar]
  8. Henriques F., Park R. B. Spectral characterization of five chlorophyll-protein complexes. Plant Physiol. 1978 Dec;62(6):856–860. doi: 10.1104/pp.62.6.856. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henriques F., Park R. Polypeptide composition of chlorophyll-protein complexes from romaine lettuce. Plant Physiol. 1977 Jul;60(1):64–68. doi: 10.1104/pp.60.1.64. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Higgins R. C., Dahmus M. E. Rapid visualization of protein bands in preparative SDS-polyacrylamide gels. Anal Biochem. 1979 Mar;93(2):257–260. doi: 10.1016/s0003-2697(79)80148-7. [DOI] [PubMed] [Google Scholar]
  11. Kirchanski S. J., Park R. B. Comparative Studies of the Thylakoid Proteins of Mesophyll and Bundle Sheath Plastids of Zea mays. Plant Physiol. 1976 Sep;58(3):345–349. doi: 10.1104/pp.58.3.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lane L. C. A simple method for stabilizing protein-sulfhydryl groups during SDS-gel electrophoresis. Anal Biochem. 1978 Jun 1;86(2):655–664. doi: 10.1016/0003-2697(78)90792-3. [DOI] [PubMed] [Google Scholar]
  13. Machold O., Meister A. Resolution of the light-harvesting chlorophyll a/b-protein of vicia faba chloroplasts into two different chlorophyll-protein complexes. Biochim Biophys Acta. 1979 Jun 5;546(3):472–480. doi: 10.1016/0005-2728(79)90082-3. [DOI] [PubMed] [Google Scholar]
  14. Morgenthaler J. J., Marsden M. P., Price C. A. Factors affecting the separation of photosynthetically competent chloroplasts in gradients of silica sols. Arch Biochem Biophys. 1975 May;168(1):289–301. doi: 10.1016/0003-9861(75)90253-2. [DOI] [PubMed] [Google Scholar]
  15. Remy R., Hoarau J., Leclerc J. C. Electrophoretic and spectrophotometric studies of chlorophyll-protein complexes from tobacco chloroplasts. Isolation of a light harvesting pigment protein complex with a molecular weight of 70,000. Photochem Photobiol. 1977 Aug;26(2):151–158. doi: 10.1111/j.1751-1097.1977.tb07466.x. [DOI] [PubMed] [Google Scholar]
  16. Strotmann H., Hesse H., Edelmann K. Quantitative determination of coupling factor CF1 of chloroplasts. Biochim Biophys Acta. 1973 Aug 31;314(2):202–210. doi: 10.1016/0005-2728(73)90135-7. [DOI] [PubMed] [Google Scholar]
  17. Wessels J. S., Borchert M. T. Polypeptide profiles of chlorophyll . protein complexes and thylakoid membranes of spinach chloroplasts. Biochim Biophys Acta. 1978 Jul 6;503(1):78–93. doi: 10.1016/0005-2728(78)90163-9. [DOI] [PubMed] [Google Scholar]

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