title: Heterologous expression of moss light-harvesting complex stress-related 1 (LHCSR1), the chlorophyll a-xanthophyll pigment-protein complex catalyzing non-photochemical quenching, in Nicotiana sp.
abstract: Oxygenic photosynthetic organisms evolved mechanisms for thermal dissipation of energy absorbed in excess to prevent formation of reactive oxygen species. The major and fastest component, called non-photochemical quenching, occurs within the photosystem II antenna system by the action of two essential light-harvesting complex (LHC)-like proteins, photosystem II subunit S (PSBS) in plants and light-harvesting complex stress-related (LHCSR) in green algae and diatoms. In the evolutionary intermediate Physcomitrella patens, a moss, both gene products are active. These proteins, which are present in low amounts, are difficult to purify, preventing structural and functional analysis. Here, we report on the overexpression of the LHCSR1 protein from P. patens in the heterologous systems Nicotiana benthamiana and Nicotiana tabacum using transient and stable nuclear transformation. We show that the protein accumulated in both heterologous systems is in its mature form, localizes in the chloroplast thylakoid membranes, and is correctly folded with chlorophyll a and xanthophylls but without chlorophyll b, an essential chromophore for plants and algal LHC proteins. Finally, we show that recombinant LHCSR1 is active in quenching in vivo, implying that the recombinant protein obtained is a good material for future structural and functional studies.
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Alberta
authors.initials: A
authors.lastname: Pinnola
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Ghin
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Gecchele
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Matilde
authors.initials: M
authors.lastname: Merlin
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Alboresi
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Linda
authors.initials: L
authors.lastname: Avesani
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Pezzotti
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Capaldi
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Cazzaniga
authors.affiliation: From the Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134 Verona, Italy roberto.bassi@univr.it.
authors.forename: Roberto
authors.initials: R
authors.lastname: Bassi
mesh_terms: D019068:Bryopsida / Q000378:metabolism*; D002734:Chlorophyll / Q000737:chemistry*; D000077194:Chlorophyll A; D008027:Light; D045342:Light-Harvesting Protein Complexes / Q000096:biosynthesis* / Q000737:chemistry; D014975:Lutein / Q000737:chemistry; D010777:Photochemistry; D010788:Photosynthesis; D045332:Photosystem II Protein Complex / Q000737:chemistry; D018515:Plant Leaves / Q000378:metabolism; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D013347:Subcellular Fractions; D020524:Thylakoids / Q000737:chemistry; D014026:Nicotiana / Q000378:metabolism*; D024341:Xanthophylls / Q000737:chemistry*
chemical_list: D045342:Light-Harvesting Protein Complexes; D045332:Photosystem II Protein Complex; D011994:Recombinant Proteins; D024341:Xanthophylls; D002734:Chlorophyll; C037184:chlorophyll b; D014975:Lutein; D000077194:Chlorophyll A
keywords: LHCSR; NPQ; Physcomitrella; light-harvesting complex (antenna complex); photobiology; photosynthesis; photosynthetic pigment; recombinant protein expression; tobacco