Cloning, expression, crystallization and preliminary X-ray studies of the ferredoxin-NAD(P)+reductase from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1.
Cloning, expression, crystallization and preliminary X-ray studies of the ferredoxin-NAD(P)+reductase from the thermophilic cyanobacterium Thermosynechococcus elongatus BP-1.
复制标题
来自嗜热蓝细菌Thermosynechochococcus elongatus BP-1的铁氧还蛋白-NAD(P)还原酶的克隆、表达、结晶和初步X射线研究。
DOI：
10.1107/s1744309112031910
复制
发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Kurisu G.
中科院分区：
文献类型：
--
作者：
Liauw P;Mashiba T;Kopczak M;Wiegand K;Muraki N;Kubota H;Kawano Y;Ikeuchi M;Hase T;Rogner M;Kurisu G.
铁氧还蛋白-NADP+ 还原酶 (FNR) 是一种黄素酶，可在光合电子传递链的最后一步中催化 NADP+ 的还原。与叶绿体 FNR 相比，来自嗜热蓝细菌 Thermosynechococochocaccus elongatus BP-1 (TeFNR) 的 FNR 在其 N 末端包含一个额外的 9 kDa 结构域，并且比来自嗜温蓝细菌的 FNR 更热稳定。为了了解TeFNR热稳定性的结构基础并将结构作用分配给小的附加结构域，编码带有和不带有附加结构域的TeFNR的基因被设计用于异源表达，并对重组蛋白进行纯化和结晶。没有附加域的TeFNR晶体属于P21空间群，晶胞参数a = 55.05，b = 71.66，c = 89.73 Å，α = 90，β = 98.21，γ = 90°。
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 (TeFNR) 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 TeFNR and assigning a structural role to the small additional domain, the gene encoding TeFNR with and without an additional domain was engineered for heterologous expression and the recombinant proteins were purified and crystallized. Crystals of TeFNR without the additional domain belonged to space group P21, with unit-cell parameters a = 55.05, b = 71.66, c = 89.73 Å, α = 90, β = 98.21, γ = 90°.