Goltermann N R, Rehfeld J F, Petersen H R
J Neurochem. 1980 Aug;35(2):479-83. doi: 10.1111/j.1471-4159.1980.tb06290.x.
The in vivo biosynthesis of cholecystokinin (CCK) in subcortical regions of the rat brain was studied by intracisternal pulse injections of [35S]methionine. The rats were killed 1.0 or 1.5 h after the injection. Subcortical CCK extracted in boiling water and acetic acid was immunoabsorbed by an antiserum specific for the COOH-terminal sequence of CCK. Gel chromatography of the absorbed CCK separated four molecular forms with elution constants (Kav) of 0.08, 0.50 (corresponding to the tritriacontapeptide amide, CCK-33), 1.10 (corresponding to the COOH-terminal octapeptide, CCK-8), and 1.40 (a component which may correspond to the COOH-terminal tetrapeptide amide). Significant incorporation of [35S]methionine occurred in the largest (Kav approximately 0.08) and octapeptide-like form only. Chasing with methionine indicated a precursor relationship between the largest form and CCK-8. The results demonstrate a substantial synthesis of CCK also in subcortical regions of the brain.
通过脑池内脉冲注射[35S]甲硫氨酸,研究了大鼠脑皮质下区域胆囊收缩素(CCK)的体内生物合成。注射后1.0或1.5小时处死大鼠。在沸水中和乙酸中提取的皮质下CCK被针对CCK羧基末端序列的抗血清免疫吸附。对吸附的CCK进行凝胶色谱分离出四种分子形式,洗脱常数(Kav)分别为0.08、0.50(对应于三十三肽酰胺,CCK-33)、1.10(对应于羧基末端八肽,CCK-8)和1.40(一种可能对应于羧基末端四肽酰胺的成分)。仅在最大的(Kav约为0.08)和类八肽形式中发生了显著的[35S]甲硫氨酸掺入。用甲硫氨酸追踪表明最大形式与CCK-8之间存在前体关系。结果表明,在脑皮质下区域也大量合成了CCK。