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Photosynthesis Research

, Volume 64, Issue 2–3, pp 233–242 | Cite as

Absorption spectra of chlorophyll a and b in Lhcb protein environment

  • Gianfelice Cinque
  • Roberta Croce
  • Roberto Bassi
Article

Abstract

The spectral forms of the two chlorophyll species in higher plant Photosystem II antenna proteins have been experimentally determined within their protein environment. Recombinant CP29 and LHC II antenna proteins missing individual chromophores were obtained by over-expression in bacteria without any changing of the primary protein sequence and in vitro reconstitution. Difference absorption spectroscopy with respect to the corresponding proteins binding the complete pigment complement yielded the spectral shape and extinction of single chlorophyll a and b. A functional relation of their absorption was given by Gaussian subband decomposition covering the entire Qx and Qy optical region together with the absolute value of the molar extinction coefficient. With respect to analogous determinations reported in the literature for organic solvents, this information is valuable for further understanding the in-protein chlorophyll excited states and excited state dynamics: in particular, for the calculation of Förster transfer rates by means of chlorophyll–chlorophyll overlap integral employing the Stepanov relation for emission and single chromophore transition energies according to the results of mutational analysis of chlorophyll binding sites [Bassi et al. (1999) Proc Natl Acad Sci USA 96: 10056–10061; Remelli et al. (1999) J Biol Chem 274: 33510–33521].

chlorophylls LHC II and CP29 spectroscopy molar extinction coefficient spectral form 
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References

  1. Bassi R, Croce R, Cugini D, and Sandonà D (1999) Mutation analysis of an higher plant antenna protein provides identification of chromophores bound into multiple sites. Proc Natl Acad Sci USA 96: 10056–10061PubMedCrossRefGoogle Scholar
  2. Borisov AY (1996) Specific features of excitation migration in photosynthesis. In Jennings RC, Zucchelli G, Ghetti F and Colombetti G (eds) Light as an Energy Source and Information Carrier in Plant Physiology. NATO ASI Series A 287, pp 31–40. Plenum Press, New YorkGoogle Scholar
  3. Cantor CR and Schimmel PR (1980) Biophysical Chemistry, W.H.Freeman and Company, New York Part II, Ch 7, 364Google Scholar
  4. Cinque G, Croce R, Holzwarth A and Bassi R (2000) Energy transfer among CP29 chlorophylls: Calculated Förster rates and experimental transient absorption at room temperature. Biophys J 79: 1706–1717PubMedGoogle Scholar
  5. Croce R, Breton J and Bassi R (1996) Conformational changes induced by phosphorylation in the CP29 subunit of Photosystem II. Biochemistry 35: 11142–11148PubMedCrossRefGoogle Scholar
  6. Croce R, Remelli R, Varotto C Breton J and Bassi R (1999) The neoxanthin binding site of the major light harvesting complex (LHC II) from higher plants. FEBS Lett 456: 1–6PubMedCrossRefGoogle Scholar
  7. Croce R Weiss S and Bassi R (1999) Carotenoid binding sites of the major Light Harvesting complex (LHC II) of higher plants. J Biol Chem 274: 29613–29623PubMedCrossRefGoogle Scholar
  8. Förster T (1948) Zwischemolekulare energiewanderung und fluoreszenz. Ann Physik 2: 55–67Google Scholar
  9. Förster T (1965) Delocalized excitation and excitation transfer. In Sinanoslu O (ed) Modern Quantum Chemistry, Part III, pp 93–137. Academic Press, New YorkGoogle Scholar
  10. Gillie UK, Small GJ and Golbeck JH (1989) Nonphotochemical hole burning of the native antenna complex of photosystem I (PS I-200). J Phys Chem 83: 1620–1627CrossRefGoogle Scholar
  11. Gilmore AM and Yamamoto HY (1991) Resolution of lutein and zeaxantind using a non-encapped likely carbon-bonded C18 HPLC column. J Chromatogr 543: 137–145CrossRefGoogle Scholar
  12. Giuffra, E, Cugini, D, Croce, R and Bassi, R (1996) Reconstitution and Pigment-binding properties of Recombinant CP29. Eur J Biochem 238: 112–120PubMedCrossRefGoogle Scholar
  13. Giuffra E, Zucchelli G, Sandonà D, Croce R, Cugini D, Garlaschi FM, Bassi R and Jennings RC (1997) Analysis of some optical properties of native and reconstituted Photosystem II antenna complex, CP29, pigment binding sites can be occupated by chlorophyll a or chlorophyll b and determine the spectral forms. Biochemistry 36: 12984–12993PubMedCrossRefGoogle Scholar
  14. Gradinaru CC, Ozdemir S, Gulen D, Van Stokkum IHM, van Grondelle R, van Amerongen H (1998) The flow of excitation energy in LHCII monomers: implications for the structural model of the major plant antenna. Biophysical J 75: 3064–3077CrossRefGoogle Scholar
  15. Gradinaru CC, Pascal A, Van Mourik F, Robert B, Horton P, Van Grondelle R, Van Amerongen H. (1999) Ultrafast evolution of the excited states in the chlorophyll a/b complex CP29 from green plants studied by energy selective pump probe spectroscopy. Biochemistry 37: 1143–1149CrossRefGoogle Scholar
  16. Gulen DR, Van Grondelle R and van Amerongen H (1997) Structural information on the light harvesting complex II of green plants that can be deciphered from polarized absorption characteristics. J Phys Chem B 101: 7256–7261CrossRefGoogle Scholar
  17. Hemelrijk PW, Kwa SLS, Van Grondelle R, Dekker JP (1992) Spectroscopic properties of LHC II, the main chlorophyll a/b protein complex from cloroplast membrane. Biochim Biophys Acta 1098: 159–166Google Scholar
  18. Hirs CHW (1967) Detection of peptides by chemical methods.Meth Enzym 11: 325–329CrossRefGoogle Scholar
  19. Hoff HJ and Amsez J (1991) Visible Absorption spectroscopy of chlorophylls. In: Scheer H (ed) Chlorophylls, Ch 4.1, pp 723. CRC Press, Boca Raton, FloridaGoogle Scholar
  20. Jean JM, Chan CK and Fleming GR (1988) Electronic energy transfer in photosynthetic bacterial reaction centers. Isr J Chem 28: 169–165Google Scholar
  21. Jennings RC, Zucchelli G, Finzi L and Garlaschi FM (1996) Spectral etherogenity and energy equilibration in higher plant photosystems. In: RC Jennings, Zucchelli G, Ghetti F and Colombetti G (eds) Light as an Energy Source and Information Carrier in Plant Physiology. NATO ASI Series A 287, pp 65–74. Plenum Press, New YorkGoogle Scholar
  22. Kühlbrandt W, Wang DN and Fujiyoshi Y (1994) Atomic model of plant light-harvesting complex by electron crystallography. Nature 367: 614–621PubMedCrossRefGoogle Scholar
  23. Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate exctinction coefficient and simultaneous equations for assaying chlorophyll a and b extracted with four different solvent. Verification of the concentration of chlorophyll standards by atomic absorption spectrometry. Biochim Biophys Acta 975: 384–394Google Scholar
  24. Remelli R, Varotto C, Sandonà D, Croce R and Bassi R (1999) Chlorophyll binding to monomeric light harvesting complex: A mutational analysis of chromophore binding residues. J Biol Chem 274: 33510–33521PubMedCrossRefGoogle Scholar
  25. Rogl H and Kühlbrandt W (1999) Mutant trimers of light-harvesting complex II exhibit altered pigment content and spectroscopic features. Biochemistry 42: 16214–16222CrossRefGoogle Scholar
  26. Sauer K, Smith JRL and Schultz AJ (1966) Dimerization of chlorophyll a, chlorophyll b and bacterial chlorophyll in solution. J Am Chem Soc B 88: 2681–2688CrossRefGoogle Scholar
  27. Seely GR and Jensen RG (1965) Effects of solvents on the spectrum of chlorophyll. Spectrochim Acta 21: 1835–1845CrossRefGoogle Scholar
  28. Shipman LL and Hausmann DL (1979) Förster transfer rate for chlorophyll a. Photochem Photobiol 29: 1163–1167Google Scholar
  29. Simonetto R, Crimi M, Sandonà D, Croce R, Cinque G, Breton J and Bassi R (1999) Orientation of chlorophyll transition moment in the Higher Plant Light harvesting complex CP29. Biochemistry 38: 12974–12983PubMedCrossRefGoogle Scholar
  30. Stepanov BI (1957) A universal relation between absorption and luminescence spectra of complex molecules. Sov Phys Dokl 2: 81–84Google Scholar
  31. Yang C, Kosemund K, Cornet C, and Paulsen H (1999) Exchange of pigment-binding amino acids in light-harvesting chlorophyll a/b protein. Biochemistry 42: 16205–16213CrossRefGoogle Scholar
  32. Zucchelli G, Dainese P, Jennings RC, Breton J, Garlaschi FM and Bassi R (1994) Gaussian decomposition of absorption and linear dicroism spectra od outer antenna complexes of Photosystem II. Biochemistry 33: 8982–8990PubMedCrossRefGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Gianfelice Cinque
    • 1
  • Roberta Croce
    • 2
  • Roberto Bassi
    • 3
  1. 1.Dipartimento Scientifico e TecnologicoUniversità degli Studi di VeronaVeronaItaly
  2. 2.Max Planck Intitute für StrahlenchemieMülheim am der RuhrGermany
  3. 3.Dipartimento Scientifico e TecnologicoUniversità degli Studi di VeronaVeronaItaly

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