IDENTIFICATION OF A MUTANT HUMAN INSULIN PREDICTED TO CONTAIN A SERINE-FOR-PHENYLALANINE SUBSTITUTION
IDENTIFICATION OF A MUTANT HUMAN INSULIN PREDICTED TO CONTAIN A SERINE-FOR-PHENYLALANINE SUBSTITUTION
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DOI：
10.1073/pnas.80.24.7390
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发表时间：
1983-01-01
期刊：
影响因子：
--
通讯作者：
TAGER, H
中科院分区：
文献类型：
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作者：
SHOELSON, S;FICKOVA, M;TAGER, H
利用从高胰岛素血症糖尿病患者血清中存在的异常胰岛素的相对 HPLC（高效液相色谱）保留以及该患者的胰岛素基因等位基因中 Mbo II 限制性位点的丢失获得的信息，最近预测突变型胰岛素在位置 B24 或 B25 处含有丝氨酸取代苯丙氨酸。人[SerB24]胰岛素和[SerB25]胰岛素是通过固相肽合成和半合成使用改进的方法制备的，该方法可以一步从最终产品中除去保护基团。在反相 HPLC 分析中，两种半合成胰岛素与正常胰岛素以及彼此之间明显分离。通过HPLC和放射免疫分析对患者的免疫亲和纯化的血清胰岛素进行分析表明，当对两者进行混合研究时，胰岛素在[SerB24]胰岛素的位置上洗脱，并与类似物共洗脱。其他研究表明，[SerB24] 胰岛素和 [SerB25] 胰岛素在刺激离体大鼠脂肪细胞葡萄糖氧化方面的活性分别约为正常胰岛素的 16% 和 0.5%。患者的异常胰岛素（洛杉矶胰岛素）是人[SerB24]胰岛素。 B24 和 B25 位氨基酸替换的异常胰岛素可能与人类糖尿病有关。
Using information gained from the relative HPLC [high performance liquid chromatography] retention of an abnormal insulin present in the serum of a hyperinsulinemic diabetic patient and the loss of an Mbo II restriction site in 1 of the patient''s insulin gene alleles, it was recently predicted that the mutant insulin contained a serine-for phenylalanine substitution at position B24 or B25. Human [SerB24]insulin and [SerB25]insulin were prepared by solid-phase peptide synthesis and semisynthesis using an improved approach whereby the protecting groups can be removed from the final product in a single step. During reversed-phase HPLC analysis, the 2 semisynthetic insulins were clearly separated from normal insulin and from each other. Analysis of the patient''s immunoaffinity-purified serum insulin by HPLC and radioimmunoassay showed that the insulin eluted at the position of [SerB24]insulin and coeluted with the analog when the 2 were studied in admixture. Additional studies showed that [SerB24]insulin and [SerB25]insulin have about 16 and 0.5% of the activity of normal insulin, respectively, in stimulating glucose oxidation by isolated rat adipocytes. The patient''s abnormal insulin (insulin Los Angeles) is human [SerB24]insulin. Abnormal insulins with amino acid replacements at both positions B24 and B25 can be associated with human diabetes.