Flatt P R, Swanston-Flatt S K, Hampton S M, Bailey C J, Marks V
Biosci Rep. 1986 Feb;6(2):193-9. doi: 10.1007/BF01115006.
Specific binding of the C-peptide of proinsulin was evaluated using a transplantable NEDH rat islet cell tumour predominantly composed of insulin-secreting B-cells. Cultured tumour B-cells exhibited greater than 90% viability assessed by trypan blue exclusion, and retained the ability to form tumours with accompanying hypoglycaemia and hyperinsulinaemia after reimplantation. During binding experiments with synthetic rat C-peptide I and iodinated tyrosylated rat C-peptide I, tumour B-cells exhibited 54 +/- 6% specific binding. Displacement of tracer increased with increasing concentrations of unlabelled rat C-peptide I (0.25-1,000 ng/ml), and the specificity of binding was substantiated by reduced displacement with human C-peptide. Scatchard analysis of specific C-peptide binding revealed a curvilinear plot with upward concavity. The demonstration of specific C-peptide binding to insulin-secreting B-cells provides evidence for a physiological role of proinsulin C-peptide.
使用一种主要由分泌胰岛素的B细胞组成的可移植NEDH大鼠胰岛细胞瘤,对胰岛素原C肽的特异性结合进行了评估。通过台盼蓝排斥法评估,培养的肿瘤B细胞存活率超过90%,并且在重新植入后保留了形成肿瘤并伴有低血糖和高胰岛素血症的能力。在用合成大鼠C肽I和碘化酪氨酰化大鼠C肽I进行结合实验期间,肿瘤B细胞表现出54±6%的特异性结合。随着未标记大鼠C肽I浓度(0.25 - 1000 ng/ml)的增加,示踪剂的置换增加,并且通过人C肽置换减少证实了结合的特异性。对特异性C肽结合的Scatchard分析显示出向上凹陷的曲线。胰岛素原C肽与分泌胰岛素的B细胞特异性结合的证明为胰岛素原C肽的生理作用提供了证据。