Watkinson A, O'Sullivan A J, Burgoyne R D, Dockray G J
MRC Secretory Control Research Group, University of Liverpool, U.K.
Peptides. 1990 May-Jun;11(3):435-41. doi: 10.1016/0196-9781(90)90039-8.
We have used an antiserum to a synthetic peptide fragment of bovine chromogranin A (ChrgA)[Tyr0] bovine ChrgA (306-313): YLSKEWEDA, together with antibodies to proenkephalin-derived peptides, to measure the release of immunoreactive peptides from nicotine-stimulated cultured bovine adrenal chromaffin cells. Over a period of 6 hr the accumulation of YLSKEWEDA immunoreactivity and Met-enkephalin Arg6Gly7Leu8 (MERGL) immunoreactivity in the medium of 10 microM nicotine-stimulated cells was shown to be biphasic; the initial phase occurred in the first 15-30 min and the second phase reached a peak after 4 hr. In contrast, catecholamine release occurred monophasically over the initial 15-30 min. Investigation of the second phase of peptide accumulation revealed that it was due in part to nicotine-evoked exocytosis and in part to extracellular processing of high molecular weight precursor proteins.
我们使用了一种针对牛嗜铬粒蛋白A（ChrgA）[Tyr0]牛ChrgA（306 - 313）合成肽片段：YLSKEWEDA的抗血清，以及针对脑啡肽原衍生肽的抗体，来检测尼古丁刺激的培养牛肾上腺嗜铬细胞中免疫反应性肽的释放。在6小时的时间段内，10微摩尔尼古丁刺激的细胞培养基中YLSKEWEDA免疫反应性和甲硫氨酸脑啡肽Arg6Gly7Leu8（MERGL）免疫反应性的积累呈双相性；初始阶段发生在最初的15 - 30分钟内，第二阶段在4小时后达到峰值。相比之下，儿茶酚胺的释放在最初的15 - 30分钟内呈单相性。对肽积累第二阶段的研究表明，这部分归因于尼古丁诱发的胞吐作用，部分归因于高分子量前体蛋白的细胞外加工。
