尿苷5'-（-D-阿卓-D-呋喃糖基焦磷酸）的合成、表征及性质

Synthesis, characterization and properties of uridine 5'-( -D-apio-D-furanosyl pyrophosphate).

作者信息

Kindel P K, Watson R R

出版信息

Biochem J. 1973 Jun;133(2):227-41. doi: 10.1042/bj1330227.

DOI:10.1042/bj1330227

PMID:4723773

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1177691/

Abstract

A method was developed for synthesizing UDP-apiose [uridine 5'-(alpha-d-apio-d-furanosyl pyrophosphate)] from UDP-glucuronic acid [uridine 5'-(alpha-d-glucopyranosyluronic acid pyrophosphate)] in 62% yield with the enzyme UDP-glucuronic acid cyclase. 2. UDP-apiose had the same mobility as uridine 5'-(alpha-d-xylopyranosyl pyrophosphate) when chromatographed on paper and when subjected to paper electrophoresis at pH5.8. When [(3)H]UDP-[U-(14)C]glucuronic acid was used as the substrate for UDP-glucuronic acid cyclase, the (3)H/(14)C ratio in the reaction product was that expected if d-apiose remained attached to the uridine. In separate experiments doubly labelled reaction product was: (a) hydrolysed at pH2 and 100 degrees C for 15min; (b) degraded at pH8.0 and 100 degrees C for 3min; (c) used as a substrate in the enzymic synthesis of [(14)C]apiin. In each type of experiment the reaction products were isolated and identified and were found to be those expected if [(3)H]UDP-[U-(14)C]apiose was the starting compound. 3. Chemical characterization established that the product containing d-[U-(14)C]apiose and phosphate formed on alkaline degradation of UDP-[U-(14)C]apiose was alpha-d-[U-(14)C]apio-d-furanosyl 1:2-cyclic phosphate. 4. Chemical characterization also established that the product containing d-[U-(14)C]apiose and phosphate formed on acid hydrolysis of alpha-d-[U-(14)C]apio-d-furanosyl 1:2-cyclic phosphate was d-[U-(14)C]apiose 2-phosphate. 5. The half-life periods for the degradation of UDP-[U-(14)C]apiose to alpha-d-[U-(14)C]apio-d-furanosyl 1:2-cyclic phosphate and UMP at pH8.0 and 80 degrees C, at pH8.0 and 25 degrees C and at pH8.0 and 4 degrees C were 31.6s, 97.2min and 16.5h respectively. The half-life period for the hydrolysis of UDP-[U-(14)C]-apiose to d-[U-(14)C]apiose and UDP at pH3.0 and 40 degrees C was 4.67min. After 20 days at pH6.2-6.6 and 4 degrees C, 17% of the starting UDP-[U-(14)C]apiose was degraded to alpha-d-[U-(14)C]apio-d-furanosyl 1:2-cyclic phosphate and UMP and 23% was hydrolysed to d-[U-(14)C]apiose and UDP. After 120 days at pH6.4 and -20 degrees C 2% of the starting UDP-[U-(14)C]apiose was degraded and 4% was hydrolysed.

摘要

开发了一种方法，可利用UDP - 葡萄糖醛酸环化酶从UDP - 葡萄糖醛酸[尿苷5' -（α - D - 吡喃葡萄糖醛酸焦磷酸）]合成UDP - 阿比糖[尿苷5' -（α - D - 阿比 - D - 呋喃糖基焦磷酸）]，产率为62%。2. UDP - 阿比糖在纸上色谱分析以及在pH5.8条件下进行纸电泳时，其迁移率与尿苷5' -（α - D - 吡喃木糖基焦磷酸）相同。当[(³H)UDP - [U - (¹⁴C)]葡萄糖醛酸用作UDP - 葡萄糖醛酸环化酶的底物时，反应产物中的³H/¹⁴C比值符合如果D - 阿比糖仍与尿苷相连时的预期值。在单独的实验中，对双重标记的反应产物进行了如下处理：(a) 在pH2和100℃下水解15分钟；(b) 在pH8.0和100℃下降解3分钟；(c) 用作[(¹⁴C)]芹菜素酶促合成的底物。在每种类型的实验中，对反应产物进行了分离和鉴定，发现其为预期的产物，前提是[(³H)UDP - [U - (¹⁴C)]阿比糖是起始化合物。3. 化学表征确定，UDP - [U - (¹⁴C)]阿比糖碱性降解后形成的含有D - [U - (¹⁴C)]阿比糖和磷酸盐的产物是α - D - [U - (¹⁴C)]阿比 - D - 呋喃糖基1:2 - 环磷酸酯。4. 化学表征还确定，α - D - [U - (¹⁴C)]阿比 - D - 呋喃糖基1:2 - 环磷酸酯酸水解后形成的含有D - [U - (¹⁴C)]阿比糖和磷酸盐的产物是D - [U - (¹⁴C)]阿比糖2 - 磷酸酯。5. 在pH8.0和80℃、pH8.0和25℃以及pH8.0和4℃条件下，UDP - [U - (¹⁴C)]阿比糖降解为α - D - [U - (¹⁴C)]阿比 - D - 呋喃糖基1:2 - 环磷酸酯和UMP的半衰期分别为31.6秒、97.2分钟和16.5小时。在pH3.0和40℃条件下，UDP - [U - (¹⁴C)] - 阿比糖水解为D - [U - (¹⁴C)]阿比糖和UDP的半衰期为4.67分钟。在pH6.2 - 6.6和4℃条件下放置20天后，起始的UDP - [U - (¹⁴C)]阿比糖中有17%降解为α - D - [U - (¹⁴C)]阿比 - D - 呋喃糖基1:2 - 环磷酸酯和UMP，23%水解为D - [U - (¹⁴C)]阿比糖和UDP。在pH6.4和 - 20℃条件下放置120天后，起始的UDP - [U - (¹⁴C)]阿比糖中有2%降解，4%水解。