title: Recombinant water-soluble chlorophyll protein from Brassica oleracea var. Botrys binds various chlorophyll derivatives.
abstract: A gene coding for water-soluble chlorophyll-binding protein (WSCP) from Brassica oleracea var. Botrys has been used to express the protein, extended by a hexahistidyl tag, in Escherichia coli. The protein has been refolded in vitro to study its pigment binding behavior. Recombinant WSCP was found to bind two chlorophylls (Chls) per tetrameric protein complex but no carotenoids in accordance with previous observations with the native protein [Satoh, H., Nakayama, K., Okada, M. (1998) J. Biol. Chem. 273, 30568-30575]. WSCP binds Chl a, Chl b, bacteriochlorophyll a, and the Zn derivative of Chl a but not pheophytin a, indicating that the central metal ion in Chl is essential for binding. WSCP also binds chlorophyllides a and b and even the more distant Chl precursor Mg-protoporphyrin IX; however, these pigments fail to induce oligomerization of the protein. We conclude that the phytol group in bound Chl plays a role in the formation of tetrameric WSCP complexes. If WSCP in fact binds Chl or its derivative(s) in vivo, the lack of carotenoids in pigmented WSCP raises the question of how photooxidation, mediated by triplet-excited Chl and singlet oxygen, is prohibited. We show by spin-trap electron-paramagnetic resonance that the light-induced singlet-oxygen formation of WSCP-bound Chl is lower by a factor of about 4 than that of unbound Chl. This as-yet-unknown mechanism of WSCP to protect its bound Chl against photooxidation supports the notion that WSCP may function as a transient carrier of Chl or its derivatives.
authors.affiliation: Institut für Allgemeine Botanik der Johannes-Gutenberg-Universität, Müllerweg 6, D-55099 Mainz, Germany.
authors.forename: Kristin
authors.initials: K
authors.lastname: Schmidt
authors.forename: Christian
authors.initials: C
authors.lastname: Fufezan
authors.forename: Anja
authors.initials: A
authors.lastname: Krieger-Liszkay
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Satoh
authors.forename: Harald
authors.initials: H
authors.lastname: Paulsen
mesh_terms: D001937:Brassica / Q000235:genetics / Q000378:metabolism*; D002338:Carotenoids / Q000378:metabolism; D002734:Chlorophyll / Q000031:analogs & derivatives* / Q000378:metabolism*; D002735:Chlorophyllides / Q000378:metabolism; D002942:Circular Dichroism; D018744:DNA, Plant / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D008027:Light; D045342:Light-Harvesting Protein Complexes; D010084:Oxidation-Reduction; D010777:Photochemistry; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D011524:Protoporphyrins / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D026082:Singlet Oxygen / Q000032:analysis / Q000737:chemistry; D012995:Solubility; D013053:Spectrophotometry / Q000379:methods; D013113:Spin Labels; D014867:Water
chemical_list: D002735:Chlorophyllides; D018744:DNA, Plant; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D011524:Protoporphyrins; D011994:Recombinant Proteins; D013113:Spin Labels; D014867:Water; D002734:Chlorophyll; C039629:magnesium protoporphyrin; D026082:Singlet Oxygen; D002338:Carotenoids