Evolution of Enzymatic Activity in the Enolase Superfamily: Structural and Mutagenic Studies of the Mechanism of the Reaction Catalyzed by o-Succinylbenzoate Synthase from Escherichia coli,

化学 立体化学 羧酸盐 活动站点 大肠杆菌 ATP合酶 催化作用 生物化学 基因
作者
Vadim A. Klenchin,Erika A. Taylor,J.A. Gerlt,Ivan Rayment
出处
期刊:Biochemistry [American Chemical Society]
卷期号:42 (49): 14427-14433 被引量:28
标识
DOI:10.1021/bi035545v
摘要

o-Succinylbenzoate synthase (OSBS) from Escherichia coli, a member of the enolase superfamily, catalyzes an exergonic dehydration reaction in the menaquinone biosynthetic pathway in which 2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate (SHCHC) is converted to 4-(2'-carboxyphenyl)-4-oxobutyrate (o-succinylbenzoate or OSB). Our previous structural studies of the Mg2+·OSB complex established that OSBS is a member of the muconate lactonizing enzyme subgroup of the superfamily: the essential Mg2+ is coordinated to carboxylate ligands at the ends of the third, fourth, and fifth β-strands of the (β/α)7β-barrel catalytic domain, and the OSB product is located between the Lys 133 at the end of the second β-strand and the Lys 235 at the end of the sixth β-strand [Thompson, T. B., Garrett, J. B., Taylor, E. A, Meganathan, R., Gerlt, J. A., and Rayment, I. (2000) Biochemistry 39, 10662−76]. Both Lys 133 and Lys 235 were separately replaced with Ala, Ser, and Arg residues; all six mutants displayed no detectable catalytic activity. The structure of the Mg2+·SHCHC complex of the K133R mutant has been solved at 1.62 Å resolution by molecular replacement starting from the structure of the Mg2+·OSB complex. This establishes the absolute configuration of SHCHC: the C1−carboxylate and the C6−OH leaving group are in a trans orientation, requiring that the dehydration proceed via a syn stereochemical course. The side chain of Arg 133 is pointed out of the active site so that it cannot function as a general base, whereas in the wild-type enzyme complexed with Mg2+·OSB, the side chain of Lys 133 is appropriately positioned to function as the only acid/base catalyst in the syn dehydration. The ε-ammonium group of Lys 235 forms a cation−π interaction with the cyclohexadienyl moiety of SHCHC, suggesting that Lys 235 also stabilizes the enediolate anion intermediate in the syn dehydration via a similar interaction.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
2秒前
5秒前
科目三应助彭栋采纳,获得10
7秒前
方文浩发布了新的文献求助10
7秒前
ding应助YWang采纳,获得10
10秒前
10秒前
林宝雯关注了科研通微信公众号
15秒前
18秒前
斯文败类应助GGBOND采纳,获得10
18秒前
星辰大海应助科研通管家采纳,获得10
18秒前
李健的小迷弟应助GGBOND采纳,获得10
18秒前
上官若男应助科研通管家采纳,获得10
18秒前
19秒前
大模型应助科研通管家采纳,获得10
19秒前
圆锥香蕉应助科研通管家采纳,获得20
19秒前
星辰大海应助科研通管家采纳,获得10
19秒前
19秒前
19秒前
Bio应助科研通管家采纳,获得30
19秒前
科研通AI5应助科研通管家采纳,获得10
19秒前
斯文败类应助科研通管家采纳,获得10
19秒前
汉堡包应助科研通管家采纳,获得10
19秒前
20秒前
23秒前
25秒前
25秒前
Dotson发布了新的文献求助10
26秒前
sinsinsin发布了新的文献求助10
27秒前
CodeCraft应助娇气的天亦采纳,获得10
28秒前
29秒前
权思远发布了新的文献求助10
29秒前
彭栋发布了新的文献求助10
29秒前
量子星尘发布了新的文献求助10
30秒前
李爱国应助收集快乐采纳,获得10
31秒前
守墓人完成签到 ,获得积分10
32秒前
33秒前
科研通AI5应助xiaoxiao采纳,获得10
36秒前
顾矜应助权思远采纳,获得10
36秒前
苯氮小羊完成签到,获得积分10
36秒前
38秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3989115
求助须知:如何正确求助?哪些是违规求助? 3531367
关于积分的说明 11253688
捐赠科研通 3269986
什么是DOI,文献DOI怎么找? 1804868
邀请新用户注册赠送积分活动 882078
科研通“疑难数据库(出版商)”最低求助积分说明 809105