Nijenhuis M, Zalm R, Burbach J P
Department of Medical Pharmacology, Rudolf Magnus Institute for Neurosciences, Utrecht University, Universiteitsweg 100, 3584 CG, Utrecht, The Netherlands.
Mol Cell Endocrinol. 2000 Sep 25;167(1-2):55-67. doi: 10.1016/s0303-7207(00)00288-4.
Over 20 mutations affecting the neurophysin moiety of the vasopressin prohormone, have been identified in families suffering from familial neurohypophysial diabetes insipidus (FNDI). Only one of these, NP87E-->stop, is located outside the central conserved domain implicated in sorting of the vasopressin prohormone. To obtain clues about the mechanism of induction of FNDI by this atypical mutant we stably expressed wild type and NP87E-->stop vasopressin prohormones in (neuro)endocrine cell lines. Metabolic labeling and immunoprecipitation demonstrated reduced processing of the mutant prohormone to neurophysin. In addition, evoked secretion of neurophysin and vasopressin was diminished, suggesting that part of the mutant is retained in another intracellular compartment than the secretory granules. Indeed, immunofluorescence demonstrated accumulation of the truncated vasopressin prohormone in the endoplasmic reticulum. We conclude that the presence of the vasopressin moiety and the central conserved core of the neurophysin domain suffices for sorting and processing, but not for efficient endoplasmic reticulum exit of the vasopressin-neurophysin molecule.
在患有家族性神经垂体性尿崩症(FNDI)的家族中,已鉴定出20多种影响血管加压素前体激素中神经垂体素部分的突变。其中只有一种,即NP87E→终止密码子,位于与血管加压素前体激素分选相关的中央保守结构域之外。为了获得有关这种非典型突变体诱导FNDI机制的线索,我们在(神经)内分泌细胞系中稳定表达了野生型和NP87E→终止密码子的血管加压素前体激素。代谢标记和免疫沉淀表明,突变体前体激素加工成神经垂体素的过程减少。此外,神经垂体素和血管加压素的诱发分泌减少,这表明部分突变体保留在不同于分泌颗粒的另一个细胞内区室中。实际上,免疫荧光显示截短的血管加压素前体激素在内质网中积累。我们得出结论,血管加压素部分的存在以及神经垂体素结构域的中央保守核心足以进行分选和加工,但不足以使血管加压素-神经垂体素分子有效地从内质网中排出。