Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase.
Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase.
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DOI：
10.1039/c5sc00164a
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发表时间：
2015-05-01
期刊：
影响因子：
8.4
通讯作者：
Micklefield J
中科院分区：
文献类型：
--
作者：
Law BJC;Struck AW;Bennett MR;Wilkinson B;Micklefield J
雷帕霉素O-甲基转移酶（RapM）的表征及其在偶联反应中的利用，与人甲硫氨酸腺苷转移酶（hMAT 2A）的改进变体，产生新的区域选择性烷基化雷帕霉素衍生物。天然产物通过S-腺苷甲硫氨酸（S-adenosyl methionine，也称为SAM）依赖性甲基转移酶的甲基化是许多生物合成途径中常见的剪裁步骤。甲基取代基的引入可以影响所产生的次级代谢产物的生物学和物理化学性质。最近，已经变得明显的是，一些α-Met依赖性甲基转移酶表现出混杂性，并将接受α-Met类似物，使转移的替代烷基。在这项研究中，我们的特点是甲基转移酶，RapM，这是参与生物合成的有效的免疫抑制剂雷帕霉素。我们已经表明，重组RapM在体外区域选择性地甲基化去甲基雷帕霉素前体的C16羟基基团，并且在接受除了C3 Met之外的替代辅因子方面是混杂的。开发了偶联酶系统，包括突变的人酶甲硫氨酸腺苷转移酶（MAT），沿着RapM，其用于制备具有交替的乙基和烯丙基醚基团的烷基化雷帕霉素衍生物（rapalogs），衍生自简单的S-乙基或S-烯丙基甲硫氨酸类似物。还有另外两种甲基转移酶RapI和RapQ提供雷帕霉素的甲基取代基。因此，使用本文所述的酶促方法，应该可以产生具有改变的性质的多种烷基化雷帕霉素类似物，这将难以通过传统的合成方法获得。
Characterisation of a rapamycin O-methyltransferase (RapM) and its utilisation in coupled reactions, with an improved variant of the human methionine adenosyl transferase (hMAT2A), results in new regioselectively alkylated rapamycin derivatives. The methylation of natural products by S-adenosyl methionine (AdoMet, also known as SAM)-dependent methyltransferase enzymes is a common tailoring step in many biosynthetic pathways. The introduction of methyl substituents can affect the biological and physicochemical properties of the secondary metabolites produced. Recently it has become apparent that some AdoMet-dependent methyltransferases exhibit promiscuity and will accept AdoMet analogues enabling the transfer of alternative alkyl groups. In this study we have characterised a methyltransferase, RapM, which is involved in the biosynthesis of the potent immunosuppressive agent rapamycin. We have shown that recombinant RapM regioselectively methylates the C16 hydroxyl group of desmethyl rapamycin precursors in vitro and is promiscuous in accepting alternative co-factors in addition to AdoMet. A coupled enzyme system was developed, including a mutant human enzyme methionine adenosyl transferase (MAT), along with RapM, which was used to prepare alkylated rapamycin derivatives (rapalogs) with alternative ethyl and allyl ether groups, derived from simple S-ethyl or S-allyl methionine analogues. There are two other methyltransferases RapI and RapQ which provide methyl substituents of rapamycin. Consequently, using the enzymatic approach described here, it should be possible to generate a diverse array of alkylated rapalogs, with altered properties, that would be difficult to obtain by traditional synthetic approaches.
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影响因子：
158.5
作者：
Krueger, Darcy A.;Care, Marguerite M.;Franz, David Neal
通讯作者：
Franz, David Neal
DOI：
10.1111/nyas.12122
发表时间：
2013-01-01
期刊：
PHARMACEUTICAL SCIENCE TO IMPROVE THE HUMAN CONDITION: PRIX GALIEN USA 2012
影响因子：
--
作者：
Lebwohl, David;Anak, Oezlem;Berg, William
通讯作者：
Berg, William
影响因子：
16.6
作者：
Gregory, MA;Petkovic, H;Sheridan, RM
通讯作者：
Sheridan, RM
影响因子：
--
作者：
LUENGO, JI;YAMASHITA, DS;HOLT, DA
通讯作者：
HOLT, DA
影响因子：
3.2
作者：
Gregory, Matthew A.;Hong, Hui;Sheridan, Rose M.
通讯作者：
Sheridan, Rose M.