Fukasawa K M, Fukasawa K, Harada M
Biochim Biophys Acta. 1978 Jul 21;535(1):161-6. doi: 10.1016/0005-2795(78)90042-9.
Dipeptidyl aminopeptidase IV was purified 350 fold from pig kidney by chromatographic procedures including affinity chromatography with conjugates of Gly-Pro linked to Sepharose 4.B. Purified enzyme existed in a dimeric form as determined by sodium dodecyl sulfate polyacrylamide-gel electrophoresis using dimethyl suberimidate (a cross-linking reagent). The molecular weight of the subunit was estimated to be 100 000 by gel filtration with 6 M guanidine hydrochloride and to be 94 000 based on analysis of N-terminal residue (dinitrophenyl-serine). The amino acid composition of the purified enzyme was also determined. The enzyme contained 18.3% of carbohydrate consisting of mannose, galactose, fucose, glucosamine and sialic acid. The enzyme desialized with sialidase was found to still possess full enzyme activity.
通过包括用与琼脂糖4B相连的甘氨酰-脯氨酸缀合物进行亲和色谱在内的色谱方法,从猪肾中纯化出了350倍的二肽基肽酶IV。通过使用辛二酸二甲酯(一种交联剂)的十二烷基硫酸钠聚丙烯酰胺凝胶电泳测定,纯化的酶以二聚体形式存在。通过在6M盐酸胍中进行凝胶过滤,亚基的分子量估计为100000,基于N端残基(二硝基苯基-丝氨酸)分析为94000。还测定了纯化酶的氨基酸组成。该酶含有18.3%的碳水化合物,由甘露糖、半乳糖、岩藻糖、葡糖胺和唾液酸组成。发现用唾液酸酶去唾液酸化的酶仍具有完全的酶活性。