Cellodextrin phosphorylase from Ruminiclostridium thermocellum: X-ray crystal structure and substrate specificity analysis.
Cellodextrin phosphorylase from Ruminiclostridium thermocellum: X-ray crystal structure and substrate specificity analysis.
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DOI：
10.1016/j.carres.2017.07.005
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发表时间：
2017-11-08
影响因子：
3.1
通讯作者：
Field RA
中科院分区：
文献类型：
--
作者：
O'Neill EC;Pergolizzi G;Stevenson CEM;Lawson DM;Nepogodiev SA;Field RA
GH 94糖苷水解酶纤维糊精磷酸化酶（CDP，EC 2.4.1.49）从短β-1 β 4-葡聚糖和α-D-葡萄糖1-磷酸产生纤维糊精低聚物。与从葡萄糖和α-D-葡萄糖1-磷酸产生纤维二糖的纤维二糖磷酸化酶（CBP）相比，CDP的生物化学特性较差。在此，我们研究了来自热纤瘤胃梭菌的重组CDP的供体和受体底物特异性，并且我们用非天然供体α-D-葡萄糖胺1-磷酸分离并纯化了葡萄糖胺加成产物至纤维二糖受体。此外，我们报告了CDP的第一个X射线晶体结构，沿着与CBPs和其他密切相关的酶的可用结构进行比较，这有助于理解区分单糖（CBP）和寡糖（CDP）受体底物所需的关键结构特征。针对各种供体和受体筛选CDP活性。GlcN-1-P作为CDP供体并产生单一周转产物。Xyl-1-P和Man-1-P是CDP的弱供体但竞争性抑制剂。在非还原端具有GlcN和Xyl的寡聚体是良好的CDP受体。解析了CDP及其与纤维四糖、磷的络合物的载脂蛋白结构。
The GH94 glycoside hydrolase cellodextrin phosphorylase (CDP, EC 2.4.1.49) produces cellodextrin oligomers from short β-1→4-glucans and α-D-glucose 1-phosphate. Compared to cellobiose phosphorylase (CBP), which produces cellobiose from glucose and α-D-glucose 1-phosphate, CDP is biochemically less well characterised. Herein, we investigate the donor and acceptor substrate specificity of recombinant CDP from Ruminiclostridium thermocellum and we isolate and characterise a glucosamine addition product to the cellobiose acceptor with the non-natural donor α-D-glucosamine 1-phosphate. In addition, we report the first X-ray crystal structure of CDP, along with comparison to the available structures from CBPs and other closely related enzymes, which contributes to understanding of the key structural features necessary to discriminate between monosaccharide (CBP) and oligosaccharide (CDP) acceptor substrates. Screening of CDP activity against a variety of donors and acceptors. GlcN-1-P acts as CDP donor and generates a single turnover product. Xyl-1-P and Man-1-P are poor donors but competitive inhibitors of CDP. Oligosaccharides with GlcN and Xyl at the non-reducing end are good CDP acceptors. The apo structure of CDP and its complex with cellotetraose and Pi have been resolved.
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影响因子：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
通讯作者：
Sternberg MJ
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
影响因子：
2.9
作者：
De Groeve, M. R. M.;Tran, G. H.;Soetaert, W.
通讯作者：
Soetaert, W.
影响因子：
3.1
作者：
Hiraishi, Masao;Igarashi, Kiyohiko;Samejima, Masahiro
通讯作者：
Samejima, Masahiro
DOI：
10.1107/s090744490903947x
发表时间：
2010-04
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Cowtan K
通讯作者：
Cowtan K