Pierik A J, Roseboom W, Happe R P, Bagley K A, Albracht S P
E. C. Slater Institute, Biochemistry, University of Amsterdam, Plantage Muidergracht 12, NL-1018 TV Amsterdam, The Netherlands.
J Biol Chem. 1999 Feb 5;274(6):3331-7. doi: 10.1074/jbc.274.6.3331.
Infrared-spectroscopic studies on the [NiFe]-hydrogenase of Chromatium vinosum-enriched in 15N or 13C, as well as chemical analyses, show that this enzyme contains three non-exchangeable, intrinsic, diatomic molecules as ligands to the active site, one carbon monoxide molecule and two cyanide groups. The results form an explanation for the three non-protein ligands to iron detected in the crystal structure of the Desulfovibrio gigas hydrogenase (Volbeda, A., Garcin, E., Piras, C., De Lacey, A. I., Fernandez, V. M., Hatchikian, E. C., Frey, M., and Fontecilla-Camps, J. C. (1996) J. Am. Chem. Soc. 118, 12989-12996) and for the low spin character of the lone ferrous iron ion observed with Mössbauer spectroscopy (Surerus, K. K., Chen, M., Van der Zwaan, W., Rusnak, F. M., Kolk, M. , Duin, E. C., Albracht, S. P. J., and Münck, E. (1994) Biochemistry 33, 4980-4993). The results do not support the notion, based upon studies of Desulfovibrio vulgaris [NiFe]-hydrogenase (Higuchi, Y., Yagi, T., and Noritake, Y. (1997) Structure 5, 1671-1680), that SO is a ligand to the active site. The occurrence of both cyanide and carbon monoxide as intrinsic constituents of a prosthetic group is unprecedented in biology.
对富含(^{15}N)或(^{13}C)的嗜酒色杆菌的[NiFe] - 氢化酶进行的红外光谱研究以及化学分析表明,该酶含有三个不可交换的、内在的双原子分子作为活性位点的配体,一个一氧化碳分子和两个氰基。这些结果解释了在巨大脱硫弧菌氢化酶的晶体结构中检测到的与铁结合的三个非蛋白质配体(Volbeda, A., Garcin, E., Piras, C., De Lacey, A. I., Fernandez, V. M., Hatchikian, E. C., Frey, M., and Fontecilla - Camps, J. C. (1996) J. Am. Chem. Soc. 118, 12989 - 12996),以及用穆斯堡尔光谱观察到的孤立亚铁离子的低自旋特性(Surerus, K. K., Chen, M., Van der Zwaan, W., Rusnak, F. M., Kolk, M., Duin, E. C., Albracht, S. P. J., and Münck, E. (1994) Biochemistry 33, 4980 - 4993)。这些结果不支持基于对普通脱硫弧菌[NiFe] - 氢化酶的研究(Higuchi, Y., Yagi, T., and Noritake, Y. (1997) Structure 5, 1671 - 1680)得出的关于(SO)是活性位点配体的观点。氰化物和一氧化碳作为辅基的内在成分同时出现,这在生物学中是前所未有的。