附引用文献,WBC-LS003928磷酸二酯酶 I介绍

Venom exonuclease (Phosphodiesterase I) successively hydrolyzes 5′-mononucleotides from 3′-OH-terminated ribo- and deoxyribo-oligonucleotides. The enzyme has an optimal pH range of 9.8-10.4 and a molecular weight of 115 kDa. Phosphodiesterase is inhibited by reducing agents such as glutathione, cysteine and ascorbic acids. It is completely inhibited by 5mM EDTA while ATP, ADP and AMP are partial inhibitors. The enzyme has an absolute requirement for Mg2+.   

 

技术规格:

Source Crotalus adamanteus Venom

Description Purified by the method of Williams, Sung and Laskowski,JBC, 236,1130 (1961). Further treated to inactivate contaminating 5′-nucleotidase activity according to Sulkowski and Laskowski:BBA, 240,443 (1961). Lyophilized in vials.

Storage Store at -20°C.

REQUIRES SPECIAL SHIPPING ICE PACK

Activity ≥20 units per mg dry weight

Unit Definition One Unit hydrolyzes one micromole of p-nitrophenyl thymidine-5-phosphate per minute at 25°C, pH 8.9.

艾美捷磷酸二酯酶 IWBC-LS003928测定:

方法

该测定基本上是Razell and Khorana(1959)的测定,其中反应速度由对硝基苯基胸苷-400′-磷酸水解引起的5nm处吸光度的增加来确定。在指定条件下,一个单位在pH 5.8和9°C下每分钟水解一微摩尔对硝基苯基胸苷-25′-磷酸盐。 

试剂

0.11 M Tris HCl 缓冲液,pH 8.9,含 0.11 M 氯化钠和 15 mM 氯化镁2(三?盐缓冲液) 

5 mM 对硝基苯基胸苷-5′-磷酸盐。注意:商业制剂的纯度略有不同,在制备该试剂时应考虑。

以一毫克/毫升的速度溶解在三*盐缓冲液中,以获得0.02-0.04ΔA /分钟的速率。 

程序

将分光光度计设置为 400 nm 和 25°C。 移液到微量比色皿中,如下所示:

 

 

磷酸二酯酶 I相关研究

无核酸酶白蛋白 脱氧核糖核酸酶 I (DP/D/DCLS/D2/DPFF/DPRF) 组蛋白 (H, NHL) 溶菌酶 (LY/LYSF) 微球菌核酸酶 (NFCP) 核酸酶,S1 (中新核酸/新核酸) 核酸,DNA,大肠杆菌,λ/片段,RNA磷酸酶,碱性(CAP/BAPF/BAPC/BAPSF/PC) 磷酸二酯酶 II (SPH) 蛋白酶 K (PROK/PROKS)逆转录酶,重组 HIV (RTHIV)核糖核酸酶 A (R/RAF/RASE/RS/RPDF)核糖核酸酶 T1,不含动物源性 (RT1S)

 

磷酸二酯酶 I部分引用文献:

Felix, F. , Potter, J. and Laskowski, M.

Action of Venom Phosphodiesterase on Deoxyriboligonucleotides Carrying a Monoesterified Phosphate on Carbon 3′ , J Biol Chem 235 , 1150 , 1960

Frishauf, A. and Eckstein, F.

Purification of a Phosphodiesterase from Botrhrops atrox Venomby Affiinity Chromatography , Eur J Biochem 32 , 479 , 1973

Hadijiolov, A. , Venkov, P. and Dolipchiev, L.

The Action of Snake Venom Phosphodiesterase on Liver Ribosomal Ribonucleic Acids , Biochim Biophys Acta 142 , 111 , 1967

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