Yagi T, Kimura K, Inokuchi H
J Biochem. 1985 Jan;97(1):181-7. doi: 10.1093/oxfordjournals.jbchem.a135042.
Hydrogenase [hydrogen: ferricytochrome c3 oxidoreductase, EC 1.12.2.1] solubilized and purified from the particulate fraction of Desulfovibrio vulgaris Miyazaki F (IAM 12604) contains 8 iron and 8 labile sulfide ions in one molecule which is composed of two unequal subunits (Mr: 60,000 + 29,000). It does not contain nickel atoms. The EPR (electron paramagnetic resonance) spectrum has an isotropic signal at g = 2.017 which is independent of the temperature. The peak-to-peak width of the signal is about 20 G. The signal intensity is nearly equivalent to 1 unpaired electron per molecule. No other signals can be detected in the field range between 2,240 and 4,240 G (which corresponds to g-values between 2.91 and 1.54). Ferricyanide has only a little effect on the shape and intensity of the EPR signal. The hydrogenase reduced under H2 is EPR silent. The Mössbauer spectrum has no hyperfine splitting at 4K. The isomer shift and quadrupole splitting at 77K are 0.38 and 0.87 mm/s, respectively. Based on these magnetic measurements, the structure of the active center of hydrogenase was suggested to be [4Fe-4S]3+ + [4Fe-4S]2+.
从普通脱硫弧菌宫崎F株（IAM 12604）的颗粒部分溶解并纯化得到的氢化酶[氢：铁细胞色素c3氧化还原酶，EC 1.12.2.1]，在由两个不等亚基（Mr：60,000 + 29,000）组成的一个分子中含有8个铁和8个不稳定硫化物离子。它不含镍原子。电子顺磁共振（EPR）谱在g = 2.017处有一个各向同性信号，该信号与温度无关。信号的峰峰宽约为20 G。信号强度几乎相当于每个分子1个未配对电子。在2,240至4,240 G的磁场范围内（对应于g值在2.91和1.54之间）未检测到其他信号。铁氰化物对EPR信号的形状和强度影响很小。在H2下还原的氢化酶EPR无信号。穆斯堡尔谱在4K时没有超精细分裂。在77K时的同质异能位移和四极分裂分别为0.38和0.87 mm/s。基于这些磁性测量，推测氢化酶活性中心的结构为[4Fe-4S]3+ + [4Fe-4S]2+。
