Open AccessCloning, expression, crystallization and preliminary X‐ray studies of the ferredoxin–NAD(P) + reductase from the thermophilic cyanobacterium Thermosynechococcus elongatus BP‐1
Author(s) -
Liauw Pasqual,
Mashiba Tomohiro,
Kopczak Marta,
Wiegand Katrin,
Muraki Norifumi,
Kubota Hisako,
Kawano Yusuke,
Ikeuchi Masahiko,
Hase Toshiharu,
Rögner Matthias,
Kurisu Genji
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112031910
Subject(s) - thermophile , thermostability , ferredoxin , biochemistry , reductase , ferredoxin—nadp(+) reductase , cyanobacteria , oxidoreductase , heterologous expression , recombinant dna , biology , chemistry , gene , enzyme , genetics , bacteria
Ferredoxin–NADP + reductase (FNR) is a flavoenzyme that catalyses the reduction of NADP + in the final step of the photosynthetic electron‐transport chain. FNR from the thermophilic cyanobacterium Thermosynechococcus elongatus BP‐1 ( Te FNR) contains an additional 9 kDa domain at its N‐terminus relative to chloroplastic FNRs and is more thermostable than those from mesophilic cyanobacteria. With the aim of understanding the structural basis of the thermostability of Te FNR and assigning a structural role to the small additional domain, the gene encoding Te FNR with and without an additional domain was engineered for heterologous expression and the recombinant proteins were purified and crystallized. Crystals of Te FNR without the additional domain belonged to space group P 2 1 , with unit‐cell parameters a = 55.05, b = 71.66, c = 89.73 Å, α = 90, β = 98.21, γ = 90°.