Iguchi H, Natori S, Kato K, Nawata H, Chrétien M
Department of Biochemistry, National Kyushu Cancer Center, Fukuoka, Japan.
Life Sci. 1988;43(23):1945-52. doi: 10.1016/s0024-3205(88)80013-4.
Chromogranin B 420-493 (GAWK)-like immunoreactivity (chromogranin B (420-493)-LI) was determined by radioimmunoassay using two different rabbit antisera, one raised against chromogranin B (420-436) (GAWK 1-17) (Ab420-436) and the other against chromogranin B 439-457 (GAWK 20-38) (Ab439-457), in bovine and human tissues. Chromogranin B (420-493)-LI was present in the bovine adrenal medulla chromaffin granules as well as in the anterior pituitary gland and was released from the cultured bovine chromaffin cells by stimulation with high K+ or nicotine. Chromogranin B (420-493)-LI present in the bovine tissues was detected using Ab420-436 but was not detected using Ab439-457. In the human tissues, chromogranin B (420-493)-LI was detected using Ab420-436 as well as Ab439-457. This suggests that the amino acid sequence of this region (chromogranin B 439-457) is different between human and bovine. On the gel permeation chromatography, chromogranin B (420-493)-LI was eluted at the void volume in the bovine adrenal medulla and at an apparent molecular weight of 4000 in the anterior pituitary gland. On the reverse-phase high-performance liquid chromatography, multiple peaks of chromogranin B (420-493)-LI was detected in the bovine adrenal medulla while one component of chromogranin B (420-493)-LI was found in the anterior pituitary gland. These results suggest that chromogranin B is processed into small fragments of chromogranin B (420-493)-LIs and that this processing is tissue-specific.
使用两种不同的兔抗血清，通过放射免疫分析法测定嗜铬粒蛋白B 420 - 493（GAWK）样免疫反应性（嗜铬粒蛋白B（420 - 493）-LI），一种抗血清针对嗜铬粒蛋白B（420 - 436）（GAWK 1 - 17）（Ab420 - 436）制备，另一种针对嗜铬粒蛋白B 439 - 457（GAWK 20 - 38）（AbS439 - 457），用于检测牛和人类组织。嗜铬粒蛋白B（420 - 493）-LI存在于牛肾上腺髓质嗜铬颗粒以及垂体前叶中，并通过高钾或尼古丁刺激从培养的牛嗜铬细胞中释放出来。使用Ab420 - 436可检测到牛组织中存在的嗜铬粒蛋白B（420 - 493）-LI，但使用Ab439 - 457未检测到。在人类组织中，使用Ab420 - 436以及Ab439 - 457均可检测到嗜铬粒蛋白B（420 - 493）-LI。这表明该区域（嗜铬粒蛋白B 439 - 457）的氨基酸序列在人和牛之间存在差异。在凝胶渗透色谱法中，嗜铬粒蛋白B（420 - 493）-LI在牛肾上腺髓质中于空体积处洗脱，在垂体前叶中表观分子量为4000处洗脱。在反相高效液相色谱法中，在牛肾上腺髓质中检测到嗜铬粒蛋白B（420 - 493）-LI的多个峰，而在垂体前叶中发现嗜铬粒蛋白B（420 - 493）-LI的一个组分。这些结果表明嗜铬粒蛋白B被加工成嗜铬粒蛋白B（420 - 493）-LI的小片段，并且这种加工具有组织特异性。
