Study on the reaction mechanism of peroxidase based on its tertiary structure

基于过氧化物酶三级结构的反应机理研究

• 批准号: 06680654
• 负责人: FUKUYAMA Keiichi
• 金额: $ 1.34万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06680654/
• 关键词: Peroxidase; X-ray crystallographic analysis; Three-dimensional structure of protein; Coordination structure of heme iron

1. Sequences of genomic DNA and cDNA coding for Arthromyces ramosus peroxidase (ARP) have been determined. The mature ARP consists of 344 amino acids with N-terminal pyroglutamic acid preceded by a signal peptide of 20 amino acid residues.2. High resolution structures of cyanide- and triiodide-bound forms of ARP have shown that on ligand binding the conformation of the distal histidine changed markedly but that of distal arginine changed little. The heme iron in the cyanide complex (low-spin ferric) is located on the heme plane, whereas the iron in the triiodide complex (high-spin ferric) is displaced from the plane about 0.2A toward the proximal side.3. Spectroscopic and crystallographic studies have shown that under physiological conditions the heme iron of ARP is in the pentacoordinated high-spin state and that at high pH the heme iron is able to bind ammonia forming the low-spin state. Comparison of the geometries around the water molecules at the distal side of the heme in peroxidases has suggested that the location of the water molecule is dominated by the orientation of the imidazole ring of the distal histidine rather than by interaction with the heme iron.4. Two proposed calcium sites have been confirmed by X-ray anomalous scattering technique.

1。基因组DNA和cDNA编码的ramosus过氧化物酶（ARP）的编码。成熟的ARP由344个氨基酸组成，其N末端硫酸谷氨酸之前是20个氨基酸残基的信号肽。2。氰化物和三碘化物结合形式的ARP的高分辨率结构表明，在配体结合远端组氨酸的构象上明显变化，但远端精氨酸的构素发生了明显变化。氰化物络合物（低旋转铁）中的血红素铁位于血红素平面上，而三碘化物复合物（高旋转铁）中的铁从平面上移动约0.2a向近端。3。光谱学和晶体学研究表明，在生理条件下，ARP的血红素铁在五型高旋转状态下，在高pH值下，血红素铁能够结合形成低旋转状态的氨。过氧化物酶末端的水分子周围几何形状的比较表明，水分子的位置由远端组氨酸的咪唑环的方向主导，而不是与血红素铁相互作用。4。通过X射线异常散射技术证实了两个提出的钙位点。

项目成果

国島直樹: "糸状菌ペルオキシダーゼの構造と機能" バイオサイエンスとインダストリー. 53. 857-863 (1995)

Naoki Kunishima：“丝状真菌过氧化物酶的结构和功能”生物科学与工业 53. 857-863 (1995)。

N. Kunishima: "Pentacoordination of the heme iron of Arthromyces ramosus peroxidase shown by a 1.8Å resolution crystallographic study at pH4.5" FEBS Letters. 378. 291-294 (1996)

N. Kunishima：“在 pH4.5 下通过 1.8Å 分辨率晶体学研究显示支枝节霉过氧化物酶的血红素铁的五配位” FEBS Letters 378. 291-294 (1996)。

N.Kunishima, F.Ameda, K.Fukuyama, M.Kawamoto, T.Matsunaga, H.Matsubara: "Pentacoordination of the heme iron of Arthromyces ramosus percxidase shown by a 1.8A resolution crystallographic study at pH 4.5" FEBS Letters. 378. 291-294 (1996)

N.Kunishima、F.Ameda、K.Fukuyama、M.Kawamoto、T.Matsunaga、H.Matsubara：“通过在 pH 4.5 下进行的 1.8A 分辨率晶体学研究显示节肢菌过氧化酶的血红素铁的五配位” FEBS Letters。

福山恵一: "ペルオキシダーゼの立体構造" 日本農芸化学会誌. 68. 1697-1700 (1994)

Keiichi Fukuyama：“过氧化物酶的立体结构”日本农业化学学会杂志 68. 1697-1700 (1994)。

N.Kunishima: "Peutocoordination of the heme iron of Arthromyces ramosus perxidase shown by a 1.8Å resolution crystallographic study at pH4.5" FEBS Letters. 378. 291-294 (1996)

N.Kunishima：“在 pH4.5 下通过 1.8Å 分辨率晶体学研究显示节枝菌过氧化物酶的血红素铁的Peutocoordination” FEBS Letters 378. 291-294 (1996)。