ENZYMIC AND IMMUNOCHEMICAL PROPERTIES OF LYSOZYME .3. EVALUATION OF SEVERAL AMINO GROUP REVERSIBLE BLOCKING REAGENTS
ENZYMIC AND IMMUNOCHEMICAL PROPERTIES OF LYSOZYME .3. EVALUATION OF SEVERAL AMINO GROUP REVERSIBLE BLOCKING REAGENTS
复制标题
DOI：
10.1021/bi00827a016
复制
发表时间：
1970-01-01
期刊：
影响因子：
2.9
通讯作者：
ATASSI, MZ
中科院分区：
文献类型：
--
作者：
HABEEB, AFSA;ATASSI, MZ
材料与方法溶菌酶（三次结晶）从Sigma Chemical Co.获得。冻干的溶壁微球菌细胞在来自Worthington Biochemical Corp.的小瓶中;马来酸酐，购自Fisher Scientific Co.;柠檬酸酐，来自Eastman Organic Chemicals; β-琥珀酸酐，来自K & K Laboratories，Inc.;来自奥尔德里奇化学公司的双乙烯酮和5，5 '-二硫代双（硝基苯甲酸）。溶菌酶与马来酸酐的反应和封闭基的去除。将酸酐作为在二氧六环中的储备溶液（50 mg/ml）加入到溶菌酶溶液（2%，在0.1M磷酸盐缓冲液中，pH 8.0）中，并且量在0.25和4摩尔试剂/游离氨基之间变化，以获得具有可变数目的氨基的制剂。反应在室温下进行30分钟，之后将蛋白质用水透析并冷冻干燥。加入的二氧六环的最大浓度
Materials and MethodsMaterials. Lysozyme (three-times crystallized) was obtained from Sigma Chemical Co. Lyophilized Micrococcus lysodeikticus cells were in vials from Worthington Biochemical Corp.; maleic anhydride from Fisher Scientific Co.; citraconic anhydride from Eastman Organic Chemicals; tetrafluoro-succinic anhydridefrom K & K Laboratories, Inc.; diketene and 5, 5'-dithiobis (nitrobenzoic acid) from Aldrich Chemical Co.Reaction of Lysozyme with Maleic Anhydride and Removal of Blocking Groups. The anhydride was added as a stock solution (50 mg/ml) in dioxane to a solution of lysozyme (2% in 0.1 m phosphate buffer, pH 8.0) and the amount varied between 0.25 and 4 mole of reagent per free amino group in order to obtain preparations with variable number of amino groups. Reaction was at room temperature for 30 min after which the proteins were dialyzed against water and freeze-dried. The maximum concentration of added dioxane