title: In vitro-mutagenesis of NADPH:protochlorophyllide oxidoreductase B: two distinctive protochlorophyllide binding sites participate in enzyme catalysis and assembly.
abstract: NADPH:protochlorophyllide oxidoreductase (POR) B is a key enzyme for the light-induced greening of etiolated angiosperm plants. It is nucleus-encoded, imported into the plastids posttranslationally, and assembled into larger light-harvesting POR:protochlorophyllide complexes termed LHPP (Reinbothe et al., Nature 397:80-84, 1999). An in vitro-mutagenesis approach was taken to study the role of the evolutionarily conserved Cys residues in pigment binding. Four Cys residues are present in the PORB of which two, Cys276 and Cys303, established distinct pigment binding sites, as shown by biochemical tests, protein import studies, and in vitro-reconstitution experiments. While Cys276 constituted the Pchlide binding site in the active site of the enzyme, Cys303 established a second, low affinity pigment binding site that was involved in the assembly and stabilization of imported PORB enzyme inside etioplasts.
authors.affiliation: Lehrstuhl für Pflanzenphysiologie, Universität Bayreuth, Universitätsstrasse 30, 95447, Bayreuth, Germany. christiane.reinbothe@uni-bayreuth.de
authors.forename: Christiane
authors.initials: C
authors.lastname: Reinbothe
authors.forename: Frank
authors.initials: F
authors.lastname: Buhr
authors.forename: Sandra
authors.initials: S
authors.lastname: Bartsch
authors.forename: Claire
authors.initials: C
authors.lastname: Desvignes
authors.forename: Françoise
authors.initials: F
authors.lastname: Quigley
authors.forename: Hélène
authors.initials: H
authors.lastname: Pesey
authors.forename: Steffen
authors.initials: S
authors.lastname: Reinbothe
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002384:Catalysis; D017931:DNA Primers; D008969:Molecular Sequence Data; D016296:Mutagenesis; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers; D044925:Oxidoreductases Acting on CH-CH Group Donors; C023886:protochlorophyllide reductase