Functional characterization of TtnD and TtnF, unveiling new insights into tautomycetin biosynthesis.
Functional characterization of TtnD and TtnF, unveiling new insights into tautomycetin biosynthesis.
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DOI：
10.1021/ja9082446
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发表时间：
2010-05-19
影响因子：
15
通讯作者：
Shen, Ben
中科院分区：
文献类型：
--
作者：
Luo, Yinggang;Li, Wenli;Ju, Jianhua;Yuan, Qiuping;Peters, Noel R.;Hoffmann, F. Michael;Huang, Sheng-Xiong;Bugni, Tim S.;Rajski, Scott;Osada, Hiroyuki;Shen, Ben
互变霉素（TTN）是一种从灰产色链霉菌中分离的高效选择性蛋白磷酸酶（PP）抑制剂，其生物合成基因簇已被克隆和测序。为了更好地理解负责将后聚酮合酶产物转化为令人兴奋的抗癌和免疫抑制化疗候选物TTN的转化，我们在S.灰发色菌ttnD和ttnF分别编码推定的脱羧酶和脱羧酶，失活得到突变株SB 13013和SB 13014。ΔttnD突变体SB 13013积累了四种新的TTN类似物TTN D-1、TTN D-2、TTN D-3和TTN D-4，而ΔttnF突变体仅积累了一种新的TTN类似物TTN F-1。这些新TTN类似物的积累定义了TtnD和TtnF的功能以及它们与TTN内C5酮部分的安装相关的化学反应的时间。值得注意的是，所有新的类似物具有结构上不同的羧酸部分，揭示TtnD显然不能催化脱羧在TtnF的情况下。此外，细胞毒性和PP抑制测定揭示了由TtnDF安装的官能团的重要性，并且与先前的提议一致，TTN的C2”-C5片段是TTN显示的重要且独特的PP-1选择性背后的关键结构决定因素。
The biosynthetic gene cluster for tautomycetin (TTN), a highly potent and selective protein phosphatase (PP) inhibitor isolated from Streptomyces griseochromogenes, has recently been cloned and sequenced. To better understand the transformations responsible for converting the post-polyketide synthase product into the exciting anticancer and immunosuppressive chemotherapeutic candidate TTN, we produced and characterized new analogues resulting from inactivation of two genes, ttnD and ttnF, in S. griseochromogenes. Inactivation of ttnD and ttnF, which encode for putative decarboxylase and dehydratase enzymes, respectively, afforded mutant strains SB13013 and SB13014. The ΔttnD mutant SB13013 accumulated four new TTN analogues, TTN D-1, TTN D-2, TTN D-3, and TTN D-4 whereas the ΔttnF mutant accumulated only one new TTN analogue, TTN F-1. The accumulation of these new TTN analogues defines the function of TtnD and TtnF and the timing of their chemistries in relation to installation of the C5 ketone moiety within TTN. Notably, all new analogues possess a structurally distinguishing carboxylic acid moiety, revealing that TtnD apparently cannot catalyze decarboxylation in the absence of TtnF. Additionally, cytotoxicity and PP inhibition assays reveal the importance of the functional groups installed by TtnDF and, consistent with earlier proposals, the C2"-C5 fragment of TTN to be a critical structural determinant behind the important and unique PP-1 selectivity displayed by TTN.
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影响因子：
5.2
作者：
Ju, Jianhua;Li, Wenli;Shen, Ben
通讯作者：
Shen, Ben
影响因子：
2.7
作者：
Liu, W;Sheppeck, JE;Chamberlin, AR
通讯作者：
Chamberlin, AR
影响因子：
15
作者：
OHTANI, I;KUSUMI, T;KAKISAWA, H
通讯作者：
KAKISAWA, H
聚酮化合物脱羧链终止之前是素硫蛋白A生物合成中的O-磺化。
影响因子：
15
作者：
Gu, Liangcai;Wang, Bo;Kulkarni, Amol;Gehret, Jennifer J.;Lloyd, Kayla R.;Gerwick, Lena;Gerwick, William H.;Wipf, Peter;Hakansson, Kristina;Smith, Janet. L.;Sherman, David H.
通讯作者：
Sherman, David H.
DOI：
10.1073/pnas.0337542100
发表时间：
2003-02-18
影响因子：
11.1
作者：
Gust, B;Challis, GL;Chater, KF
通讯作者：
Chater, KF