Crystal structure of a phenol-coupling P450 monooxygenase involved in teicoplanin biosynthesis.
Crystal structure of a phenol-coupling P450 monooxygenase involved in teicoplanin biosynthesis.
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DOI：
10.1002/prot.22996
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发表时间：
2011-06
影响因子：
2.9
通讯作者：
Nair, Satish K.
中科院分区：
文献类型：
--
作者：
Li, Zhi;Rupasinghe, Sanjeewa G.;Schuler, Mary A.;Nair, Satish K.
脂糖肽类抗生素替考拉宁已被证明对革兰氏阳性病原体有效。替考拉宁与万古霉素型糖肽类抗生素的区别在于芳香族氨基酸1和3之间存在额外的交联，该交联由细胞色素P450单加氧酶Orf 6 *（CYP 165 D3）催化。作为一个目标，对了解这种苯酚偶联反应的机制，我们已经确定了重组Orf 6 * 和确定其晶体结构，以2.2 μ m分辨率。虽然Orf 6 * 的结构揭示了其他P450单加氧酶共有的核心折叠，但在容纳其复杂七肽底物所需的活性位点空腔附近的二级结构元件的配置中存在细微差异。具体而言，Orf 6 * 中F和G螺旋的取向导致比万古霉素氧化酶OxyB和OxyC中发现的活性位点更封闭的活性位点。此外，Met 226在I螺旋取代更典型的甘氨酸/丙氨酸残基，位于血红素卟啉环上方，在那里它与血红素铁结合的水分子形成氢键。与其他酚偶联P450单加氧酶的序列比较表明，Met 226在确定Orf 6 * 的底物区域特异性中起作用。这些特征为糖肽类抗生素生物合成过程中发生的交联机制提供了进一步的见解。
The lipoglycopeptide antibiotic teicoplanin has proven efficacy against gram-positive pathogens. Teicoplanin is distinguished from the vancomycin-type glycopeptide antibiotics, by the presence of an additional cross-link between the aromatic amino acids 1 and 3 that is catalyzed by the cytochrome P450 monooxygenase Orf6* (CYP165D3). As a goal towards understanding the mechanism of this phenol-coupling reaction, we have characterized recombinant Orf6* and determined its crystal structure to 2.2 Å resolution. Although the structure of Orf6* reveals the core fold common to other P450 monooxygenases, there are subtle differences in the disposition of secondary structure elements near the active site cavity necessary to accommodate its complex heptapeptide substrate. Specifically, the orientation of the F and G helices in Orf6* results in a more closed active site than found in the vancomycin oxidative enzymes OxyB and OxyC. In addition, Met226 in the I helix replaces the more typical Gly/Ala residue that is positioned above the heme porphyrin ring, where it forms a hydrogen bond with a heme iron-bound water molecule. Sequence comparisons with other phenol-coupling P450 monooxygenases suggest that Met226 plays a role in determining the substrate regiospecificity of Orf6*. These features provide further insights into the mechanism of the cross-linking mechanisms that occur during glycopeptide antibiotics biosynthesis.
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DOI：
10.1073/pnas.0805983105
发表时间：
2008-10-14
影响因子：
11.1
作者：
Cryle, Max J.;Schlichting, Ilme
通讯作者：
Schlichting, Ilme
DOI：
10.1107/s0907444903018043
发表时间：
2003-11-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
Grosse-Kunstleve, RW;Adams, PD
通讯作者：
Adams, PD
影响因子：
3.3
作者：
MALABARBA, A;FERRARI, P;CAVALLERI, B
通讯作者：
CAVALLERI, B
DOI：
10.1107/s0907444902016657
发表时间：
2002-11-01
影响因子：
2.2
作者：
Adams, PD;Grosse-Kunstleve, RW;Terwilliger, TC
通讯作者：
Terwilliger, TC
影响因子：
5.2
作者：
Malabarba, A;Goldstein, BP
通讯作者：
Goldstein, BP