Ito M, Oiso Y, Murase T, Kondo K, Saito H, Chinzei T, Racchi M, Lively M O
First Department of Internal Medicine, Nagoya University School of Medicine, Aichi, Japan.
J Clin Invest. 1993 Jun;91(6):2565-71. doi: 10.1172/JCI116494.
A transition of G to A at nucleotide position 279 in exon 1 of the vasopressin gene has been identified in patients with familial central diabetes insipidus. The mutation predicts an amino acid substitution of Thr (ACG) for Ala (GCG) at the COOH terminus of the signal peptide in preprovasopression (preproVP). Translation in vitro of wild-type and mutant mRNAs produced 19-kD preproVPs. When translated in the presence of canine pancreatic rough microsomes, wild-type preproVP was converted to a 21-kD protein, whereas the mutant mRNA produced proteins of 21 kD and 23 kD. NH2-terminal amino acid sequence analysis revealed that the 21-kD proteins from the wild-type and the mutants were proVPs generated by the proteolytic cleavage of the 19-residue signal peptide and the addition of carbohydrate. Accordingly, mutant preproVP was cleaved at the correct site after Thr-19, but the efficiency of cleavage by signal peptidase was < 25% that observed for the wild-type preproVP, resulting in the formation of a predominant glycosylated but uncleaved 23-kD product. These data suggest that inefficient processing of preproVP produced by the mutant allele is possibly involved in the pathogenesis of diabetes insipidus in the affected individuals.
在家族性中枢性尿崩症患者中,已鉴定出血管加压素基因外显子1中核苷酸位置279处的G到A转换。该突变预测在血管加压素原(preproVP)信号肽的COOH末端,苏氨酸(ACG)被丙氨酸(GCG)取代。野生型和突变型mRNA的体外翻译产生了19-kD的血管加压素原。当在犬胰腺粗面微粒体存在下进行翻译时,野生型血管加压素原被转化为21-kD的蛋白质,而突变型mRNA产生了21 kD和23 kD的蛋白质。NH2末端氨基酸序列分析表明,野生型和突变型的21-kD蛋白质是由19个残基信号肽的蛋白水解切割和碳水化合物添加产生的血管加压素原。因此,突变型血管加压素原在苏氨酸-19之后的正确位点被切割,但信号肽酶的切割效率不到野生型血管加压素原的25%,导致形成主要的糖基化但未切割的23-kD产物。这些数据表明,突变等位基因产生的血管加压素原加工效率低下可能与受影响个体的尿崩症发病机制有关。