Uechi Keiko, Takata Goro, Yoneda Kazunari, Ohshima Toshihisa, Sakuraba Haruhiko
Rare Sugar Research Center, Kagawa University, 2393 Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0701, Japan.
Department of Bioscience, School of Agriculture, Tokai University, Aso, Kumamoto 869-1404, Japan.
Acta Crystallogr F Struct Biol Commun. 2014 Jul;70(Pt 7):890-5. doi: 10.1107/S2053230X14011005. Epub 2014 Jun 18.
The crystal structure of a D-tagatose 3-epimerase-like protein (MJ1311p) encoded by a hypothetical open reading frame, MJ1311, in the genome of the hyperthermophilic archaeon Methanocaldococcus jannaschii was determined at a resolution of 2.64 Å. The asymmetric unit contained two homologous subunits, and the dimer was generated by twofold symmetry. The overall fold of the subunit proved to be similar to those of the D-tagatose 3-epimerase from Pseudomonas cichorii and the D-psicose 3-epimerases from Agrobacterium tumefaciens and Clostridium cellulolyticum. However, the situation at the subunit-subunit interface differed substantially from that in D-tagatose 3-epimerase family enzymes. In MJ1311p, Glu125, Leu126 and Trp127 from one subunit were found to be located over the metal-ion-binding site of the other subunit and appeared to contribute to the active site, narrowing the substrate-binding cleft. Moreover, the nine residues comprising a trinuclear zinc centre in endonuclease IV were found to be strictly conserved in MJ1311p, although a distinct groove involved in DNA binding was not present. These findings indicate that the active-site architecture of MJ1311p is quite unique and is substantially different from those of D-tagatose 3-epimerase family enzymes and endonuclease IV.
对嗜热古菌詹氏甲烷球菌基因组中一个假定的开放阅读框MJ1311编码的D-塔格糖3-表异构酶样蛋白(MJ1311p)的晶体结构进行了测定,分辨率为2.64 Å。不对称单元包含两个同源亚基,二聚体通过二重对称产生。亚基的整体折叠结构被证明与来自菊苣假单胞菌的D-塔格糖3-表异构酶以及来自根癌农杆菌和溶纤梭菌的D-阿洛酮糖3-表异构酶相似。然而,亚基-亚基界面的情况与D-塔格糖3-表异构酶家族酶有很大不同。在MJ1311p中,发现一个亚基的Glu125、Leu126和Trp127位于另一个亚基的金属离子结合位点上方,似乎对活性位点有贡献,使底物结合裂隙变窄。此外,尽管不存在参与DNA结合的明显凹槽,但发现内切核酸酶IV中构成三核锌中心的九个残基在MJ1311p中严格保守。这些发现表明,MJ1311p的活性位点结构非常独特,与D-塔格糖3-表异构酶家族酶和内切核酸酶IV的活性位点结构有很大不同。