Structure of the Photoreactive Iron Center of the Nitrile Hydratase from Rhodococcus sp. N-771
Structure of the Photoreactive Iron Center of the Nitrile Hydratase from Rhodococcus sp. N-771
复制标题
红球菌腈水合酶光反应铁中心的结构。
DOI：
--
复制
发表时间：
1997
影响因子：
4.8
通讯作者：
I. Endo
中科院分区：
文献类型：
--
作者：
M. Tsujimura;N. Dohmae;M. Odaka;M. Chijimatsu;K. Takio;M. Yohda;M. Hoshino;S. Nagashima;I. Endo
腈水合酶（Nitrile hydratase，NHase）是一种光反应性酶，由非血红素铁中心的亚硝基化失活，一氧化氮（nitric oxide，NO）的光解激活。为了获得铁中心的结构信息，我们通过蛋白水解分离含有铁中心的肽复合物。当通过反相高效液相色谱法分析从非活性形式分离的α亚基的胰蛋白酶消化物时，在肽片段Asn 105-Val-Ile-Val-Cys-Ser-Leu-Cys-Ser-Cys-Thr-Ala-Trp-Pro-Ile-Leu-Gly-Leu-Pro-Pro-Thr-Trp-Tyr-Lys 128中观察到亚硝基化铁中心的吸光度特征。该肽含有0.79 mol铁/mol分子以及内源性NO。随后，通过用嗜热菌蛋白酶、羧肽酶Y和亮氨酸氨肽酶M消化该肽，我们发现亚硝基化铁中心所需的最小肽段是从α Ile 107到α Trp 117的11个氨基酸残基。此外，通过使用质谱、蛋白质序列和氨基酸组成分析，我们已经表明α亚基的第112个Cys残基在NH酶中被后氧化为半胱氨酸-亚磺酸（Cys-SO2 H）。这些结果表明，来自红球菌属物种N-771的腈水合酶具有在铁中心含有半胱氨酸-亚磺酸的新型非血红素铁酶。可能的铁中心的配体残基进行了讨论。
Nitrile hydratase (NHase) fromRhodococcus sp. N-771 is a photoreactive enzyme that is inactivated by nitrosylation of the non-heme iron center and activated by photodissociation of nitric oxide (NO). To obtain structural information on the iron center, we isolated peptide complexes containing the iron center by proteolysis. When the tryptic digest of the α subunit isolated from the inactive form was analyzed by reversed-phase high performance liquid chromatography, the absorbance characteristic of the nitrosylated iron center was observed in the peptide fragment, Asn105-Val-Ile-Val-Cys-Ser-Leu-Cys-Ser-Cys-Thr-Ala-Trp-Pro-Ile-Leu-Gly-Leu-Pro-Pro-Thr-Trp-Tyr-Lys128. The peptide contained 0.79 mol of iron/mol of molecule as well as endogenous NO. Subsequently, by digesting the peptide with thermolysin, carboxypeptidase Y, and leucine aminopeptidase M, we found that the minimum peptide segment required for the nitrosylated iron center is the 11 amino acid residues from αIle107 to αTrp117. Furthermore, by using mass spectrometry, protein sequence, and amino acid composition analyses, we have shown that the 112th Cys residue of the α subunit is post-translationally oxidized to a cysteine-sulfinic acid (Cys-SO2H) in the NHase. These results indicate that the NHase from Rhodococcus sp. N-771 has a novel non-heme iron enzyme containing a cysteine-sulfinic acid in the iron center. Possible ligand residues of the iron center are discussed.
以 2.8 A 分辨率精制肠球菌 NADH 过氧化物酶的天然半胱氨酸-磺酸氧化还原中心的结构。
DOI：
10.1021/bi961037s
发表时间：
1996
期刊：
Biochemistry.
影响因子：
--
作者：
Yeh,JI;Claiborne,A;Hol,WG
通讯作者：
Hol,WG
影响因子：
56.9
作者：
STORZ, G;TARTAGLIA, LA;AMES, BN
通讯作者：
AMES, BN