Kinetic, mutational, and structural analysis of malonate semialdehyde decarboxylase from Coryneform bacterium strain FG41: mechanistic implications for the decarboxylase and hydratase activities.
Kinetic, mutational, and structural analysis of malonate semialdehyde decarboxylase from Coryneform bacterium strain FG41: mechanistic implications for the decarboxylase and hydratase activities.
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DOI：
10.1021/bi400567a
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发表时间：
2013-07-16
期刊：
影响因子：
2.9
通讯作者：
Whitman, Christian P.
中科院分区：
文献类型：
--
作者：
Guo, Youzhong;Serrano, Hector;Poelarends, Gerrit J.;Johnson, William H., Jr.;Hackert, Marvin L.;Whitman, Christian P.
来自 Pseudomonas pavonaceae 170 的丙二酸半醛脱羧酶（称为 Pp MSAD）位于杀线虫剂 1,3-二氯丙烯的细菌分解代谢途径中。 MSAD 具有两种已知的活性：它催化丙二酸半醛不依赖于金属离子的脱羧生成乙醛和二氧化碳，以及催化 2-氧代-3-戊炔酸的低水平水合生成乙酰丙酮酸。目前尚不清楚后一种活性是否具有生物学相关性。先前的研究确定 Pro-1、Asp-37 和一对精氨酸（Arg-73 和 Arg-75）是这些活性中的关键残留物。来自棒状杆菌菌株 FG41 的 MSAD（指定为 FG41 MSAD）与假单胞菌酶（包括 Pro-1 和 Asp-37）具有 38% 的成对序列同一性。然而，Gln-73 取代了 Arg-73，并且第二个精氨酸通过插入甘氨酸而转变为 Arg-76。为了确定这些变化与 FG41 MSAD 活性的关系，克隆了该基因，并对酶进行了表达和表征。该酶具有相当的脱羧酶活性，但水合酶活性显着降低。诱变以及天然酶（2.0 Å 分辨率）和 3-氧代丙酸酯部分修饰的酶（由酶和 3-溴丙炔酯孵育产生）（2.2 Å 分辨率）的晶体结构提供了结构基础。 Pro-1 和 Asp-37 的作用可能与 MSAD 的作用相同。但Thr-72、Gln-73、Tyr-123的侧链在机制上取代了Arg-73、Arg-75的侧链，发挥结合和催化作用。这些结构还表明，Arg-76 可能距离太远，无法在该机制中发挥直接作用。 FG41 MSAD 是互变异构酶超家族 MSAD 家族中第二个功能注释的同源物，可能代表一个新的亚家族。
Malonate semialdehyde decarboxylase from Pseudomonas pavonaceae 170 (designated Pp MSAD) is in a bacterial catabolic pathway for the nematicide 1,3-dichloropropene. MSAD has two known activities: it catalyzes the metal-ion independent decarboxylation of malonate semialdehyde to produce acetaldehyde and carbon dioxide, as well as a low-level hydration of 2-oxo-3-pentynoate to yield acetopyruvate. The latter activity is not known to be biologically relevant. Previous studies identified Pro-1, Asp-37, and a pair of arginines (Arg-73 and Arg-75) as critical residues in these activities. MSAD from Coryneform bacterium strain FG41 (designated FG41 MSAD) shares 38% pairwise sequence identity with the Pseudomonas enzyme including Pro-1 and Asp-37. However, Gln-73 replaces Arg-73, and the second arginine is shifted to Arg-76 by the insertion of a glycine. In order to determine how these changes relate to the activities of FG41 MSAD, the gene was cloned and the enzyme expressed and characterized. The enzyme has a comparable decarboxylase activity, but a significantly reduced hydratase activity. Mutagenesis along with crystal structures of the native enzyme (2.0 Å resolution) and the enzyme modified by a 3-oxopropanoate moiety (resulting from the incubation of enzyme and 3-bromopropiolate) (2.2 Å resolution) provided a structural basis. The roles of Pro-1 and Asp-37 are likely the same as those proposed for MSAD. However, the side chains of Thr-72, Gln-73, and Tyr-123 replace those of Arg-73 and Arg-75 in the mechanism and play a role in binding and catalysis. The structures also show that Arg-76 is likely too distant to play a direct role in the mechanism. FG41 MSAD is the second functionally annotated homologue in the MSAD family of the tautomerase superfamily and could represent a new subfamily.
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影响因子：
2.9
作者：
Burks, Elizabeth A.;Fleming, Christopher D.;Mesecar, Andrew D.;Whitman, Christian P.;Pegan, Scott D.
通讯作者：
Pegan, Scott D.
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
DOI：
10.1099/00221287-137-8-2025
发表时间：
1991-08-01
期刊：
JOURNAL OF GENERAL MICROBIOLOGY
影响因子：
--
作者：
HARTMANS, S;JANSEN, MW;DEBONT, JAM
通讯作者：
DEBONT, JAM
影响因子：
2.9
作者：
Almrud, JJ;Poelarends, GJ;Whitman, CP
通讯作者：
Whitman, CP
DOI：
10.1107/s0907444998003254
发表时间：
1998-09-01
期刊：
ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY
影响因子：
--
作者：
Brunger, AT;Adams, PD;Warren, GL
通讯作者：
Warren, GL