Fermani Simona, Sparla Francesca, Marri Lucia, Thumiger Anton, Pupillo Paolo, Falini Giuseppe, Trost Paolo
Department of Chemistry, University of Bologna, Via Selmi 2, 40126 Bologna, Italy.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Jun 1;66(Pt 6):621-6. doi: 10.1107/S1744309110013527. Epub 2010 May 25.
The crystal structure of the A(4) isoform of photosynthetic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from Arabidopsis thaliana, expressed in recombinant form and complexed with NAD, is reported. The crystals, which were grown in 2.4 M ammonium sulfate and 0.1 M sodium citrate, belonged to space group I222. The asymmetric unit includes ten subunits, i.e. two independent tetramers plus a dimer that generates a third tetramer by a crystallographic symmetry operation. The crystal structure was solved by molecular replacement and refined to an R factor of 23.7% and an R(free) factor of 28.9% at 2.6 A resolution. In the final model, each subunit binds one NAD(+) molecule and two sulfates, which occupy the P(s) and the P(i) anion-binding sites. Detailed knowledge of this structure is instrumental for structural investigation of supramolecular complexes of A(4)-GAPDH, phosphoribulokinase and CP12, which are involved in the regulation of photosynthesis in the model plant A. thaliana.
报道了拟南芥光合甘油醛-3-磷酸脱氢酶(GAPDH)A(4)同工型的晶体结构,该同工型以重组形式表达并与NAD复合。晶体在2.4 M硫酸铵和0.1 M柠檬酸钠中生长,属于空间群I222。不对称单元包含十个亚基,即两个独立的四聚体加上一个二聚体,该二聚体通过晶体学对称操作产生第三个四聚体。晶体结构通过分子置换法解析,并在2.6 Å分辨率下精修至R因子为23.7%,R(free)因子为28.9%。在最终模型中,每个亚基结合一个NAD(+)分子和两个硫酸盐,它们占据P(s)和P(i)阴离子结合位点。对该结构的详细了解有助于对A(4)-GAPDH、磷酸核酮糖激酶和CP12的超分子复合物进行结构研究,这些复合物参与了模式植物拟南芥的光合作用调控。