Udp-Glucose → UDP GalN biotin

HRID 882685

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HRID 882685 반응식
[CH:1]1[C:7](=[O:8])[NH:6][C:4](=[O:5])[N:3]([C@@H:9]2[O:13][C@H:12]([CH2:14][O:15][P:16]([O:19][P:20]([O:23][C@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@H]3O)([OH:22])=[O:21])([OH:18])=[O:17])[C@@H:11]([OH:35])[C@H:10]2[OH:36])[CH:2]=1.C1C(=O)NC(=[O:41])N([C@@H]2O[C@H](COP(OP([O:59][C@H:60]3[O:65][C@H:64]([CH2:66][OH:67])[C@H:63]([OH:68])[C@H:62]([OH:69])[C@H:61]3[NH2:70])(O)=O)(O)=O)[C@@H](O)[C@H]2O)C=1.S(C(N1C(=O)CCC1=O)(CCCCN[C:86](=[O:100])[CH2:87][CH2:88][CH2:89][CH2:90][C@H:91]1[C@@H:99]2[C@@H:94]([NH:95][C:96]([NH:98]2)=[O:97])[CH2:93][S:92]1)C([O-])=O)(O)(=O)=O>>[CH:1]1[C:7](=[O:8])[NH:6][C:4](=[O:5])[N:3]([C@@H:9]2[O:13][C@H:12]([CH2:14][O:15][P:16]([O:19][P:20]([OH:22])([OH:23])=[O:21])([OH:18])=[O:17])[C@@H:11]([OH:35])[C@H:10]2[OH:36])[CH:2]=1.[OH:59][CH:60]1[O:65][C@H:64]([CH2:66][OH:67])[C@H:63]([OH:68])[C@H:62]([OH:69])[C@H:61]1[NH2:70].[OH:41][C:86]([CH2:87][CH2:88][CH2:89][CH2:90][C@H:91]1[C@@H:99]2[C@@H:94]([NH:95][C:96]([NH:98]2)=[O:97])[CH2:93][S:92]1)=[O:100]

참가 물질

4

반응물 3

HCID 8629Udp-GlucoseHCID 161225UDP-galactosamineHCID 1317298646-[5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-2-(2,5-dioxopyrrolidin-1-yl)-2-sulfohexanoate

생성물 1

HCID 131729863UDP GalN biotin

실험 절차

UDP-galactosamine (UDP-GalN) is formed from UDP-Glc and galactosamine-1-phosphate by the action of galactose-1-phosphate uridyltransferase (E.C. 2.7.7.12; Sigma, USA). A typical synthesis protocol is described below. The reaction mixture contains 10 mM galactosamine-1-phosphate, 20 mM UDP-Glc, 5 U/ml of galactose-1-phosphate uridyltransferase, 100 mM Na-HEPES pH 8.0, 5 mM MgCl2, and 5 mM β-mercaptoethanol. The reaction vessel is incubated at room temperature under nitrogen atmosphere for 3 days, after which nucleotide sugars are isolated from the reaction mixture with a graphitised carbon column essentially as in (Mäki et al., 2002). The nucleotide sugar mixture, containing UDP-Glc and UDP-GalN, is incubated with a molar excess of sulfosuccinimidyl-6-(biotinamido)hexanoate (sulfo-NHS-LC-biotin; Pierce, USA) in 50 mM NH4HCO3 at room temperature for 2.5 hours. The product, UDP-GalN-biotin, uridine 5′-diphospho-N-(6-biotinamidohexanoyl)galactosamine, is purified by gel filtration and reversed phase HPLC.

후처리

  1. customat room temperature
  2. customfor 2.5 hours
  3. filtrationThe product, UDP-GalN-biotin, uridine 5′-diphospho-N-(6-biotinamidohexanoyl)galactosamine, is purified by gel filtration

출처 및 근거

ORD 기록
ord-3619111ce1b54746beae6e022093081f
출처 데이터셋
uspto-grants-2012_08
특허 번호
US08236487B2

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