PYRUVATE FORMATE-LYASE MECHANISM INVOLVING THE PROTEIN-BASED GLYCYL RADICAL
PYRUVATE FORMATE-LYASE MECHANISM INVOLVING THE PROTEIN-BASED GLYCYL RADICAL
复制标题
DOI：
10.1042/bst0210731
复制
发表时间：
1993-08-01
影响因子：
3.9
通讯作者：
WAGNER, AFV
中科院分区：
文献类型：
--
作者：
KNAPPE, J;ELBERT, S;WAGNER, AFV
伊里亚伊。提夫；提奥里。在1‘1~1中，次亚磷酸盐11I，闭合；在旋转性，提供了直接的Evi-dt，NC。蛋白质-自由基在关键的第一反应中的作用。在CC键的合成过程中，通过；In；ILOG：UC~rexts Nitti the Acetyl-R‘Interiediate生成1-羟乙基膦酸根。以(磷酰基)甲酯1 10 I.1的形式与Cys‘LS结合，形成该末端产物(结构为S11)。图2b中的n)是；1c。由于失去了甘氨酸根而引起的。检测到一种中间体；通过EPR进行了鉴定。以α-膦酰基c；II-Boii为中心的i-tiydrosyc的rxhxl的光谱研究。他的部首是rem；irk：ible st；il>；lr；it Oy‘(Ti 20 Rnin)。另一种有效的PITI灭活剂是THC。丙酮酸~iixilog~i~；icet~lplios~~liiii~ite11 11 1，它们有！C~h；Ir；黄化以产生硅自由基内部。我们有一个中心！‘PERT’或寡核苷酸-ctiwctd来研究ahovt。Cys‘“在正常的1’121反应中的所有意义。1‘1；1，iiirtmts 3 18，1 10或7.31位的标准取代，用i？表示。克隆了染色体p$‘和uct基因的大肠埃希菌，发现分离的非自由基核糖氨酸的结构完整性被正常酶的限制性蛋白水解谱所证实。’L‘He(；iy’-Alii mut；irt不能转化为；i r；iic；il-c。ONT；LILLING形式(它与
Iriae. tiv; itiori. of 1'1~ 1, hy hypophosphite 1 1 I, a close; in; ilogue of torinate, has provided direct evi-dt, nc. e tor protein-radical p; irticipation in the crucial first hnlf-reac% on. hlimicking CC bond synthesis, the; in; ilog: uc~ rexts nitti the acetyl-R'interniediate to produce 1-hydroxyethyl phosphonate that, rern; irk; itdy. is cov; iIentIy hound to Cys' lS as a (phosplioryl) tliioester 1 10 I. 1; ormation of this deadend product (structure s1iou. n in Figure 2b) is; ic. coiiip; inied by loss of the glycq'l radical. An interiiicdiate has been detected; ind identified through epr. spectronwtric studies as the a-phosphonyl c.; ii-boii-centred rxhxl of I-tiydrosyc. thyl phos-phmitv.'I'his suhstrate-b; ised radical is rem; irk: ibly st; il> lr; it OY'(ti 20 rnin). A further potent niec~ li; iiiisiii-l~~ ise~~ inactivator of PITI, is thc. pyruv-ate~ iixilog~ i~; icet~ lplios~~ liiii~ ite 1 1 1 1, which\ve have recciitl!. c~ h; ir; icterized to prodrice the siiiiie radical interinc. di; itr 12 1.Lye have rccentl!'pert'orrned oligon~ icleotide-ctiwcttd inutageiiesis to investigate ahovt. all the signiticmce of Cys'" in the nornial 1'121, reaction. 1'1; 1, iiiirtmts bvitli standard substitutions in the 3 18, 1 10 or 7.31 positions\z'ere expressed in I?. coli cells that I; ic-ked the chromosomal p $'and uct genes,; ind the structiml integrities of isolated non-radical riizyine fornis\vere verified by the limited proteolysis pattern chiracterized for the normal enzyme.'l'he (; Iy'"-Alii mut; irit was not convertible to; I r; idic; il-c. ont; iiIling form (it hiid no iiftiinity with the