Wang J, Takeuchi T, Tanaka S, Kubo S K, Kayo T, Lu D, Takata K, Koizumi A, Izumi T
Department of Molecular Medicine, Gunma University, Maebashi, Japan.
J Clin Invest. 1999 Jan;103(1):27-37. doi: 10.1172/JCI4431.
The mouse autosomal dominant mutation Mody develops hyperglycemia with notable pancreatic beta-cell dysfunction. This study demonstrates that one of the alleles of the gene for insulin 2 in Mody mice encodes a protein product that substitutes tyrosine for cysteine at the seventh amino acid of the A chain in its mature form. This mutation disrupts a disulfide bond between the A and B chains and can induce a drastic conformational change of this molecule. Although there was no gross defect in the transcription from the wild-type insulin 2 allele or two alleles of insulin 1, levels of proinsulin and insulin were profoundly diminished in the beta cells of Mody mice, suggesting that the number of wild-type (pro)insulin molecules was also decreased. Electron microscopy revealed a dramatic reduction of secretory granules and a remarkably enlarged lumen of the endoplasmic reticulum. Little proinsulin was processed to insulin, but high molecular weight forms of proinsulin existed with concomitant overexpression of BiP, a molecular chaperone in the endoplasmic reticulum. Furthermore, mutant proinsulin expressed in Chinese hamster ovary cells was inefficiently secreted, and its intracellular fraction formed complexes with BiP and was eventually degraded. These findings indicate that mutant proinsulin was trapped and accumulated in the endoplasmic reticulum, which could induce beta-cell dysfunction and account for the dominant phenotype of this mutation.
小鼠常染色体显性突变Mody会出现高血糖,并伴有明显的胰腺β细胞功能障碍。本研究表明,Mody小鼠中胰岛素2基因的一个等位基因编码的蛋白质产物,在其成熟形式的A链第七个氨基酸处,用酪氨酸替代了半胱氨酸。这种突变破坏了A链和B链之间的二硫键,并能诱导该分子发生剧烈的构象变化。虽然野生型胰岛素2等位基因或胰岛素1的两个等位基因的转录没有明显缺陷,但Mody小鼠β细胞中胰岛素原和胰岛素的水平却显著降低,这表明野生型(前)胰岛素分子的数量也减少了。电子显微镜显示分泌颗粒显著减少,内质网腔明显扩大。很少有胰岛素原加工成胰岛素,但存在高分子量形式的胰岛素原,同时内质网中的分子伴侣BiP过表达。此外,在中国仓鼠卵巢细胞中表达的突变胰岛素原分泌效率低下,其细胞内部分与BiP形成复合物并最终被降解。这些发现表明,突变胰岛素原被困在内质网中并积累,这可能导致β细胞功能障碍,并解释了这种突变的显性表型。