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. 1998 Mar 16;17(6):1577–1587. doi: 10.1093/emboj/17.6.1577

A novel multi-functional chloroplast protein: identification of a 40 kDa immunophilin-like protein located in the thylakoid lumen.

H Fulgosi 1, A V Vener 1, L Altschmied 1, R G Herrmann 1, B Andersson 1
PMCID: PMC1170505  PMID: 9501079

Abstract

We describe the identification of the first immunophilin associated with the photosynthetic membrane of chloroplasts. This complex 40 kDa immunophilin, designated TLP40 (thylakoid lumen PPIase), located in the lumen of the thylakoids, was found to play a dual role in photosynthesis involving both biogenesis and intraorganelle signalling. It originates in a single-copy nuclear gene, is made as a precursor of 49.2 kDa with a bipartite lumenal targeting transit peptide, and is characterized by a structure including a cyclophilin-like C-terminal segment of 20 kDa, a predicted N-terminal leucine zipper and a potential phosphatase-binding domain. It can exist in different oligomeric conformations and attach to the inner membrane surface. It is confined predominantly to the non-appressed thylakoid regions, the site of protein integration into the photosynthetic membrane. The isolated protein possesses peptidyl-prolyl cis-trans isomerase protein folding activity characteristic of immunophilins, but is not inhibited by cyclosporin A. TLP40 also exerts an effect on dephosphorylation of several key proteins of photosystem II, probably as a constituent of a transmembrane signal transduction chain. This first evidence for a direct role of immunophilins in a photoautotrophic process suggests that light-mediated protein phosphorylation in photosynthetic membranes and the role of the thylakoid lumen are substantially more complex than anticipated.

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Selected References

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  1. Abraham R. T. Phosphatidylinositol 3-kinase related kinases. Curr Opin Immunol. 1996 Jun;8(3):412–418. doi: 10.1016/s0952-7915(96)80132-4. [DOI] [PubMed] [Google Scholar]
  2. Adam Z. Protein stability and degradation in chloroplasts. Plant Mol Biol. 1996 Dec;32(5):773–783. doi: 10.1007/BF00020476. [DOI] [PubMed] [Google Scholar]
  3. Aldape R. A., Futer O., DeCenzo M. T., Jarrett B. P., Murcko M. A., Livingston D. J. Charged surface residues of FKBP12 participate in formation of the FKBP12-FK506-calcineurin complex. J Biol Chem. 1992 Aug 15;267(23):16029–16032. [PubMed] [Google Scholar]
  4. Allen J. F. Protein phosphorylation in regulation of photosynthesis. Biochim Biophys Acta. 1992 Jan 22;1098(3):275–335. doi: 10.1016/s0005-2728(09)91014-3. [DOI] [PubMed] [Google Scholar]
  5. Andersson B., Akerlund H. E., Albertsson P. A. Separation of subchloroplast membrane particles by counter-current distribution. Biochim Biophys Acta. 1976 Jan 15;423(1):122–132. doi: 10.1016/0005-2728(76)90106-7. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Aro E. M., Virgin I., Andersson B. Photoinhibition of Photosystem II. Inactivation, protein damage and turnover. Biochim Biophys Acta. 1993 Jul 5;1143(2):113–134. doi: 10.1016/0005-2728(93)90134-2. [DOI] [PubMed] [Google Scholar]
  9. Bierer B. E., Somers P. K., Wandless T. J., Burakoff S. J., Schreiber S. L. Probing immunosuppressant action with a nonnatural immunophilin ligand. Science. 1990 Oct 26;250(4980):556–559. doi: 10.1126/science.1700475. [DOI] [PubMed] [Google Scholar]
  10. Bose S., Weikl T., Bügl H., Buchner J. Chaperone function of Hsp90-associated proteins. Science. 1996 Dec 6;274(5293):1715–1717. doi: 10.1126/science.274.5293.1715. [DOI] [PubMed] [Google Scholar]
  11. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  12. Brandts J. F., Halvorson H. R., Brennan M. Consideration of the Possibility that the slow step in protein denaturation reactions is due to cis-trans isomerism of proline residues. Biochemistry. 1975 Nov 4;14(22):4953–4963. doi: 10.1021/bi00693a026. [DOI] [PubMed] [Google Scholar]
  13. Bugos R. C., Yamamoto H. Y. Molecular cloning of violaxanthin de-epoxidase from romaine lettuce and expression in Escherichia coli. Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6320–6325. doi: 10.1073/pnas.93.13.6320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Cardenas M. E., Hemenway C., Muir R. S., Ye R., Fiorentino D., Heitman J. Immunophilins interact with calcineurin in the absence of exogenous immunosuppressive ligands. EMBO J. 1994 Dec 15;13(24):5944–5957. doi: 10.1002/j.1460-2075.1994.tb06940.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Clausmeyer S., Klösgen R. B., Herrmann R. G. Protein import into chloroplasts. The hydrophilic lumenal proteins exhibit unexpected import and sorting specificities in spite of structurally conserved transit peptides. J Biol Chem. 1993 Jul 5;268(19):13869–13876. [PubMed] [Google Scholar]
  17. Cohen Y., Yalovsky S., Nechushtai R. Integration and assembly of photosynthetic protein complexes in chloroplast thylakoid membranes. Biochim Biophys Acta. 1995 May 8;1241(1):1–30. doi: 10.1016/0304-4157(94)00012-3. [DOI] [PubMed] [Google Scholar]
  18. Duina A. A., Chang H. C., Marsh J. A., Lindquist S., Gaber R. F. A cyclophilin function in Hsp90-dependent signal transduction. Science. 1996 Dec 6;274(5293):1713–1715. doi: 10.1126/science.274.5293.1713. [DOI] [PubMed] [Google Scholar]
  19. Freeman B. C., Toft D. O., Morimoto R. I. Molecular chaperone machines: chaperone activities of the cyclophilin Cyp-40 and the steroid aporeceptor-associated protein p23. Science. 1996 Dec 6;274(5293):1718–1720. doi: 10.1126/science.274.5293.1718. [DOI] [PubMed] [Google Scholar]
  20. Freskgård P. O., Bergenhem N., Jonsson B. H., Svensson M., Carlsson U. Isomerase and chaperone activity of prolyl isomerase in the folding of carbonic anhydrase. Science. 1992 Oct 16;258(5081):466–468. doi: 10.1126/science.1357751. [DOI] [PubMed] [Google Scholar]
  21. Fruman D. A., Burakoff S. J., Bierer B. E. Immunophilins in protein folding and immunosuppression. FASEB J. 1994 Apr 1;8(6):391–400. doi: 10.1096/fasebj.8.6.7513288. [DOI] [PubMed] [Google Scholar]
  22. Hani J., Stumpf G., Domdey H. PTF1 encodes an essential protein in Saccharomyces cerevisiae, which shows strong homology with a new putative family of PPIases. FEBS Lett. 1995 May 29;365(2-3):198–202. doi: 10.1016/0014-5793(95)00471-k. [DOI] [PubMed] [Google Scholar]
  23. Harbury P. B., Zhang T., Kim P. S., Alber T. A switch between two-, three-, and four-stranded coiled coils in GCN4 leucine zipper mutants. Science. 1993 Nov 26;262(5138):1401–1407. doi: 10.1126/science.8248779. [DOI] [PubMed] [Google Scholar]
  24. Harrison R. K., Stein R. L. Substrate specificities of the peptidyl prolyl cis-trans isomerase activities of cyclophilin and FK-506 binding protein: evidence for the existence of a family of distinct enzymes. Biochemistry. 1990 Apr 24;29(16):3813–3816. doi: 10.1021/bi00468a001. [DOI] [PubMed] [Google Scholar]
  25. Hurt E., Hauska G. A cytochrome f/b6 complex of five polypeptides with plastoquinol-plastocyanin-oxidoreductase activity from spinach chloroplasts. Eur J Biochem. 1981 Jul;117(3):591–595. doi: 10.1111/j.1432-1033.1981.tb06379.x. [DOI] [PubMed] [Google Scholar]
  26. Kadenbach B., Jarausch J., Hartmann R., Merle P. Separation of mammalian cytochrome c oxidase into 13 polypeptides by a sodium dodecyl sulfate-gel electrophoretic procedure. Anal Biochem. 1983 Mar;129(2):517–521. doi: 10.1016/0003-2697(83)90586-9. [DOI] [PubMed] [Google Scholar]
  27. Kaneko T., Sato S., Kotani H., Tanaka A., Asamizu E., Nakamura Y., Miyajima N., Hirosawa M., Sugiura M., Sasamoto S. Sequence analysis of the genome of the unicellular cyanobacterium Synechocystis sp. strain PCC6803. II. Sequence determination of the entire genome and assignment of potential protein-coding regions (supplement). DNA Res. 1996 Jun 30;3(3):185–209. doi: 10.1093/dnares/3.3.185. [DOI] [PubMed] [Google Scholar]
  28. Kennedy S., Smyth J. A., Cush P. F., McCullough S. J., Allan G. M., McQuaid S. Transatlantic spread of seal virus. Nature. 1989 Feb 23;337(6209):695–695. doi: 10.1038/337695b0. [DOI] [PubMed] [Google Scholar]
  29. Klappa P., Freedman R. B., Zimmermann R. Protein disulphide isomerase and a lumenal cyclophilin-type peptidyl prolyl cis-trans isomerase are in transient contact with secretory proteins during late stages of translocation. Eur J Biochem. 1995 Sep 15;232(3):755–764. [PubMed] [Google Scholar]
  30. Kofron J. L., Kuzmic P., Kishore V., Colón-Bonilla E., Rich D. H. Determination of kinetic constants for peptidyl prolyl cis-trans isomerases by an improved spectrophotometric assay. Biochemistry. 1991 Jun 25;30(25):6127–6134. doi: 10.1021/bi00239a007. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Lang K., Schmid F. X., Fischer G. Catalysis of protein folding by prolyl isomerase. Nature. 1987 Sep 17;329(6136):268–270. doi: 10.1038/329268a0. [DOI] [PubMed] [Google Scholar]
  33. Lippuner V., Chou I. T., Scott S. V., Ettinger W. F., Theg S. M., Gasser C. S. Cloning and characterization of chloroplast and cytosolic forms of cyclophilin from Arabidopsis thaliana. J Biol Chem. 1994 Mar 18;269(11):7863–7868. [PubMed] [Google Scholar]
  34. Liu J., Walsh C. T. Peptidyl-prolyl cis-trans-isomerase from Escherichia coli: a periplasmic homolog of cyclophilin that is not inhibited by cyclosporin A. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4028–4032. doi: 10.1073/pnas.87.11.4028. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Lu K. P., Hanes S. D., Hunter T. A human peptidyl-prolyl isomerase essential for regulation of mitosis. Nature. 1996 Apr 11;380(6574):544–547. doi: 10.1038/380544a0. [DOI] [PubMed] [Google Scholar]
  36. Luan S., Albers M. W., Schreiber S. L. Light-regulated, tissue-specific immunophilins in a higher plant. Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):984–988. doi: 10.1073/pnas.91.3.984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Luan S., Kudla J., Gruissem W., Schreiber S. L. Molecular characterization of a FKBP-type immunophilin from higher plants. Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):6964–6969. doi: 10.1073/pnas.93.14.6964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Luan S., Lane W. S., Schreiber S. L. pCyP B: a chloroplast-localized, heat shock-responsive cyclophilin from fava bean. Plant Cell. 1994 Jun;6(6):885–892. doi: 10.1105/tpc.6.6.885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Luban J., Bossolt K. L., Franke E. K., Kalpana G. V., Goff S. P. Human immunodeficiency virus type 1 Gag protein binds to cyclophilins A and B. Cell. 1993 Jun 18;73(6):1067–1078. doi: 10.1016/0092-8674(93)90637-6. [DOI] [PubMed] [Google Scholar]
  40. Lupas A. Coiled coils: new structures and new functions. Trends Biochem Sci. 1996 Oct;21(10):375–382. [PubMed] [Google Scholar]
  41. Lütcke H. A., Chow K. C., Mickel F. S., Moss K. A., Kern H. F., Scheele G. A. Selection of AUG initiation codons differs in plants and animals. EMBO J. 1987 Jan;6(1):43–48. doi: 10.1002/j.1460-2075.1987.tb04716.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Marin E., Nussaume L., Quesada A., Gonneau M., Sotta B., Hugueney P., Frey A., Marion-Poll A. Molecular identification of zeaxanthin epoxidase of Nicotiana plumbaginifolia, a gene involved in abscisic acid biosynthesis and corresponding to the ABA locus of Arabidopsis thaliana. EMBO J. 1996 May 15;15(10):2331–2342. [PMC free article] [PubMed] [Google Scholar]
  43. Marks A. R. Cellular functions of immunophilins. Physiol Rev. 1996 Jul;76(3):631–649. doi: 10.1152/physrev.1996.76.3.631. [DOI] [PubMed] [Google Scholar]
  44. Matouschek A., Rospert S., Schmid K., Glick B. S., Schatz G. Cyclophilin catalyzes protein folding in yeast mitochondria. Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6319–6323. doi: 10.1073/pnas.92.14.6319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  46. Mi H., Kops O., Zimmermann E., Jäschke A., Tropschug M. A nuclear RNA-binding cyclophilin in human T cells. FEBS Lett. 1996 Dec 2;398(2-3):201–205. doi: 10.1016/s0014-5793(96)01248-3. [DOI] [PubMed] [Google Scholar]
  47. Montague J. W., Hughes F. M., Jr, Cidlowski J. A. Native recombinant cyclophilins A, B, and C degrade DNA independently of peptidylprolyl cis-trans-isomerase activity. Potential roles of cyclophilins in apoptosis. J Biol Chem. 1997 Mar 7;272(10):6677–6684. doi: 10.1074/jbc.272.10.6677. [DOI] [PubMed] [Google Scholar]
  48. Pratt W. B. The role of heat shock proteins in regulating the function, folding, and trafficking of the glucocorticoid receptor. J Biol Chem. 1993 Oct 15;268(29):21455–21458. [PubMed] [Google Scholar]
  49. Pötter E., Kloppstech K. Effects of light stress on the expression of early light-inducible proteins in barley. Eur J Biochem. 1993 Jun 15;214(3):779–786. doi: 10.1111/j.1432-1033.1993.tb17980.x. [DOI] [PubMed] [Google Scholar]
  50. Rahfeld J. U., Rücknagel K. P., Schelbert B., Ludwig B., Hacker J., Mann K., Fischer G. Confirmation of the existence of a third family among peptidyl-prolyl cis/trans isomerases. Amino acid sequence and recombinant production of parvulin. FEBS Lett. 1994 Sep 26;352(2):180–184. doi: 10.1016/0014-5793(94)00932-5. [DOI] [PubMed] [Google Scholar]
  51. Rassow J., Mohrs K., Koidl S., Barthelmess I. B., Pfanner N., Tropschug M. Cyclophilin 20 is involved in mitochondrial protein folding in cooperation with molecular chaperones Hsp70 and Hsp60. Mol Cell Biol. 1995 May;15(5):2654–2662. doi: 10.1128/mcb.15.5.2654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Ratajczak T., Carrello A. Cyclophilin 40 (CyP-40), mapping of its hsp90 binding domain and evidence that FKBP52 competes with CyP-40 for hsp90 binding. J Biol Chem. 1996 Feb 9;271(6):2961–2965. doi: 10.1074/jbc.271.6.2961. [DOI] [PubMed] [Google Scholar]
  53. Rudd K. E., Sofia H. J., Koonin E. V., Plunkett G., 3rd, Lazar S., Rouviere P. E. A new family of peptidyl-prolyl isomerases. Trends Biochem Sci. 1995 Jan;20(1):12–14. doi: 10.1016/s0968-0004(00)88940-9. [DOI] [PubMed] [Google Scholar]
  54. Schatz G., Dobberstein B. Common principles of protein translocation across membranes. Science. 1996 Mar 15;271(5255):1519–1526. doi: 10.1126/science.271.5255.1519. [DOI] [PubMed] [Google Scholar]
  55. Schlicher T., Soll J. Molecular chaperones are present in the thylakoid lumen of pea chloroplasts. FEBS Lett. 1996 Feb 5;379(3):302–304. doi: 10.1016/0014-5793(95)01534-5. [DOI] [PubMed] [Google Scholar]
  56. Schmid F. X., Baldwin R. L. Acid catalysis of the formation of the slow-folding species of RNase A: evidence that the reaction is proline isomerization. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4764–4768. doi: 10.1073/pnas.75.10.4764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Schreiber S. L. Chemistry and biology of the immunophilins and their immunosuppressive ligands. Science. 1991 Jan 18;251(4991):283–287. doi: 10.1126/science.1702904. [DOI] [PubMed] [Google Scholar]
  58. Schreiber S. L. Immunophilin-sensitive protein phosphatase action in cell signaling pathways. Cell. 1992 Aug 7;70(3):365–368. doi: 10.1016/0092-8674(92)90158-9. [DOI] [PubMed] [Google Scholar]
  59. Sheldon P. S., Venis M. A. Purification and characterization of cytosolic and microsomal cyclophilins from maize (Zea mays). Biochem J. 1996 May 1;315(Pt 3):965–970. doi: 10.1042/bj3150965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Sokolenko A., Altschmied L., Herrmann R. G. Sodium Dodecyl Sulfate-Stable Proteases in Chloroplasts. Plant Physiol. 1997 Oct;115(2):827–832. doi: 10.1104/pp.115.2.827. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Sokolenko A., Fulgosi H., Gal A., Altschmied L., Ohad I., Herrmann R. G. The 64 kDa polypeptide of spinach may not be the LHCII kinase, but a lumen-located polyphenol oxidase. FEBS Lett. 1995 Sep 4;371(2):176–180. doi: 10.1016/0014-5793(95)00892-d. [DOI] [PubMed] [Google Scholar]
  62. Takeya T., Feldman R. A., Hanafusa H. DNA sequence of the viral and cellular src gene of chickens. 1. Complete nucleotide sequence of an EcoRI fragment of recovered avian sarcoma virus which codes for gp37 and pp60src. J Virol. 1982 Oct;44(1):1–11. doi: 10.1128/jvi.44.1.1-11.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Vener A. V., van Kan PJ, Rich P. R., Ohad I., Andersson B. Plastoquinol at the quinol oxidation site of reduced cytochrome bf mediates signal transduction between light and protein phosphorylation: thylakoid protein kinase deactivation by a single-turnover flash. Proc Natl Acad Sci U S A. 1997 Feb 18;94(4):1585–1590. doi: 10.1073/pnas.94.4.1585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  64. Walsh C. T., Zydowsky L. D., McKeon F. D. Cyclosporin A, the cyclophilin class of peptidylprolyl isomerases, and blockade of T cell signal transduction. J Biol Chem. 1992 Jul 5;267(19):13115–13118. [PubMed] [Google Scholar]
  65. von Heijne G., Steppuhn J., Herrmann R. G. Domain structure of mitochondrial and chloroplast targeting peptides. Eur J Biochem. 1989 Apr 1;180(3):535–545. doi: 10.1111/j.1432-1033.1989.tb14679.x. [DOI] [PubMed] [Google Scholar]

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