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. 1989 Apr 1;108(4):1397–1405. doi: 10.1083/jcb.108.4.1397

Lateral distribution and diffusion of plastocyanin in chloroplast thylakoids

PMCID: PMC2115525  PMID: 2647767

Abstract

The lateral distribution of plastocyanin in the thylakoid lumen of spinach and pea chloroplasts was studied by combining immunocytochemical localization and kinetic measurements of P700+ reduction at high time resolution. In dark-adapted chloroplasts, the concentration of plastocyanin in the photosystem I containing stroma membranes exceeds that in photosystem II containing grana membranes by a factor of about two. Under these conditions, the reduction of P700+ with a halftime of 12 microseconds after a laser flash of saturating intensity indicates that to greater than 95% of total photosystem I a plastocyanin molecule is bound. An analysis of the labeling densities, the length of the different lumenal regions, and the total amounts of plastocyanin and P700 shows that most of the remaining presumable mobile plastocyanin is found in the granal lumen. This distribution of plastocyanin is consistent with a more negative surface charge density in the stromal than in the granal lumen. During illumination the concentration of plastocyanin in grana increases at the expense of that in stroma lamellae, indicating a light-driven diffusion from stroma to grana regions. Our observations provide evidence that a high concentration of plastocyanin in grana in the light favors the lateral electron transport from cytochrome b6/f complexes in appressed grana across the long distance to photosystem I in nonappressed stroma membranes.

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Selected References

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  1. Allred D. R., Staehelin L. A. Implications of cytochrome b6/f location for thylakoidal electron transport. J Bioenerg Biomembr. 1986 Oct;18(5):419–436. doi: 10.1007/BF00743013. [DOI] [PubMed] [Google Scholar]
  2. Allred D. R., Staehelin L. A. Lateral Distribution of the Cytochrome b(6)/f and Coupling Factor ATP Synthetase Complexes of Chloroplast Thylakoid Membranes. Plant Physiol. 1985 May;78(1):199–202. doi: 10.1104/pp.78.1.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Allred D. R., Staehelin L. A. Spatial organization of the cytochrome b6-f complex within chloroplast thylakoid membranes. Biochim Biophys Acta. 1986 Apr 2;849(1):94–103. doi: 10.1016/0005-2728(86)90100-3. [DOI] [PubMed] [Google Scholar]
  4. Andersson B., Anderson J. M. Lateral heterogeneity in the distribution of chlorophyll-protein complexes of the thylakoid membranes of spinach chloroplasts. Biochim Biophys Acta. 1980 Dec 3;593(2):427–440. doi: 10.1016/0005-2728(80)90078-x. [DOI] [PubMed] [Google Scholar]
  5. Cox R. P., Andersson B. Lateral and transverse organisation of cytochromes in the chloroplast thylakoid membrane. Biochem Biophys Res Commun. 1981 Dec 31;103(4):1336–1342. doi: 10.1016/0006-291x(81)90269-2. [DOI] [PubMed] [Google Scholar]
  6. Gershoni J. M., Davis F. E., Palade G. E. Protein blotting in uniform or gradient electric fields. Anal Biochem. 1985 Jan;144(1):32–40. doi: 10.1016/0003-2697(85)90080-6. [DOI] [PubMed] [Google Scholar]
  7. Graan T., Ort D. R. Quantitation of the rapid electron donors to P700, the functional plastoquinone pool, and the ratio of the photosystems in spinach chloroplasts. J Biol Chem. 1984 Nov 25;259(22):14003–14010. [PubMed] [Google Scholar]
  8. Haehnel W., Pröpper A., Krause H. Evidence for complexed plastocyanin as the immediate electron donor of P-700. Biochim Biophys Acta. 1980 Dec 3;593(2):384–399. doi: 10.1016/0005-2728(80)90075-4. [DOI] [PubMed] [Google Scholar]
  9. Hardt H., Kok B. Plastocyanin as the possible site of photosynthetic electron transport inhibition by glutaraldehyde. Plant Physiol. 1977 Aug;60(2):225–229. doi: 10.1104/pp.60.2.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hardt H., Kok B. Stabilization by glutaraldehyde of high-rate electron transport in isolated chloroplasts. Biochim Biophys Acta. 1976 Oct 13;449(1):125–135. doi: 10.1016/0005-2728(76)90012-8. [DOI] [PubMed] [Google Scholar]
  11. Itoh S. Surface potential and reaction of the membrane-bound electron transfer components. II. Integrity of the chloroplast membrane and reaction of P-700. Biochim Biophys Acta. 1979 Dec 6;548(3):596–607. doi: 10.1016/0005-2728(79)90067-7. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lam E., Malkin R. Interactions between thylakoid electron transfer complexes. II. Modification studies with glutaraldehyde. Arch Biochem Biophys. 1985 Oct;242(1):64–71. doi: 10.1016/0003-9861(85)90480-1. [DOI] [PubMed] [Google Scholar]
  14. Laurell C. B. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem. 1966 Apr;15(1):45–52. doi: 10.1016/0003-2697(66)90246-6. [DOI] [PubMed] [Google Scholar]
  15. Miller K. R., Staehelin L. A. Analysis of the thylakoid outer surface. Coupling factor is limited to unstacked membrane regions. J Cell Biol. 1976 Jan;68(1):30–47. doi: 10.1083/jcb.68.1.30. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Miller T. J., Stone H. O. The rapid isolation of ribonuclease-free immunoglobulin G by protein A-sepharose affinity chromatography. J Immunol Methods. 1978;24(1-2):111–125. doi: 10.1016/0022-1759(78)90092-3. [DOI] [PubMed] [Google Scholar]
  17. Packer L., Allen J. M., Starks M. Light-induced ion tranpsort in glutaraldehyde-fixed chloroplasts: studies with nigericin. Arch Biochem Biophys. 1968 Oct;128(1):142–152. doi: 10.1016/0003-9861(68)90017-9. [DOI] [PubMed] [Google Scholar]
  18. Plesnicar M., Bendall D. S. The plastocyanin content of chloroplasts from some higher plants estimated by a sensitive enzymatic assay. Biochim Biophys Acta. 1970 Aug 4;216(1):192–199. doi: 10.1016/0005-2728(70)90170-2. [DOI] [PubMed] [Google Scholar]
  19. Rumberg B., Siggel U. pH changes in the inner phase of the thylakoids during photosynthesis. Naturwissenschaften. 1969 Mar;56(3):130–132. doi: 10.1007/BF00601025. [DOI] [PubMed] [Google Scholar]
  20. Surek B., Latzko E. Visualization of antigenic proteins blotted onto nitrocellulose using the immuno-gold-staining (IGS) method. Biochem Biophys Res Commun. 1984 May 31;121(1):284–289. doi: 10.1016/0006-291x(84)90720-4. [DOI] [PubMed] [Google Scholar]
  21. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]

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