从胎牛胰腺制备β细胞：胰岛素生物合成活性的表征

Preparation of beta-cells from fetal bovine pancreas: characterization of insulin biosynthetic activity.

作者信息

Rosenzweig S A, Yip C C

出版信息

Can J Biochem. 1979 Jun;57(6):480-8. doi: 10.1139/o79-061.

Abstract

Dispersed cell preparations enriched in beta-cells were obtained by collagenase digestion of fetal bovine pancreas and separation by Ficoll gradient centrifugation. These cells actively incorporated [3H]leucine into proinsulin and insulin. Incubation of these cells in the presence of the arginine analogue, L-canavanine, resulted in the inhibition of conversion of newly formed proinsulin to insulin and the appearance of a radioactive component of molecular weight 11,000-12,000. Incorporation of [35S]methionine into this component was detected in the presence of canavanine, an event not observed in control incubation. Canavanine thus induced the formation of a component possessing molecular weight and compositional properties expected for preproinsulin. Further characterization of cellular products by polyacrylamide slab gel electrophoresis in sodium dodecyl sulfate showed a highly labeled band corresponding to molecular weight 18,000-20,000 which might be involved in insulin biosynthesis.

摘要

通过用胶原酶消化胎牛胰腺并经Ficoll梯度离心分离，获得了富含β细胞的分散细胞制剂。这些细胞能将[3H]亮氨酸主动掺入胰岛素原和胰岛素中。在精氨酸类似物L-刀豆氨酸存在的情况下培养这些细胞，会抑制新形成的胰岛素原向胰岛素的转化，并出现分子量为11,000 - 12,000的放射性组分。在刀豆氨酸存在的情况下检测到[35S]甲硫氨酸掺入该组分，这在对照培养中未观察到。因此，刀豆氨酸诱导形成了一种具有前胰岛素原预期分子量和组成特性的组分。通过十二烷基硫酸钠聚丙烯酰胺平板凝胶电泳对细胞产物进行的进一步表征显示，有一条高度标记的条带，其分子量对应于18,000 - 20,000，这可能参与胰岛素的生物合成。