论文部分内容阅读
用NR/R双向对角线SDS-聚丙烯酰胺凝胶电泳结果表明氨基酰化酶分子中并不存在二硫键.MNP滴定氨基酰化酶的结果表明,酶分子中的10个半胱氨酸巯基有8个是可反应基团,另外2个内埋巯基在7mol/L盐酸胍溶液中被暴露出来,并可与MNP反应.DTNB测定的结果表明,酶分子中的10个半胱氨酸巯基有4个可反应巯基,6个DTNB所不能接触的巯基经7mol/L盐酸胍变性后,可以与DTNB作用.两种巯基试剂测定的结果均表明氨基酰化酶分子中含总巯基数为10个.这一数据与氨基酰化酶序列分析所得到的半胱氨酸残基数是一致的.因此进一步证实了氨基酰化酶分子中确无二硫键的存在.
SDS / polyacrylamide gel electrophoresis with NR / R bidirectional diagonal SDS-polyacrylamide gel electrophoresis showed no disulfide bond in the aminoacylase molecule.The results of MNP titration of aminoacylase showed that 10 cystemes in the enzyme molecule 8 mercapto groups were reactive groups and the other 2 mercapto groups were exposed in 7mol / L guanidine hydrochloride solution and reacted with MNP.The results of DTNB assay showed that 10 cystemes Sulfhydryl group has 4 reactive thiol groups, 6 DTNB can not be contacted with the thiol group denatured by 7mol / L guanidine hydrochloride, with DTNB role.The two sulfhydryl reagent results show that amino acylase molecules containing total thiol number Is 10. This data is consistent with the number of cysteine residues obtained from aminoacylase sequence analysis, thus further confirming the presence of a non-disulfide bond in the aminoacylase molecule.