Brand S J, Klarlund J, Schwartz T W, Rehfeld J F
J Biol Chem. 1984 Nov 10;259(21):13246-52.
Biosynthetic incorporation of labeled amino acids into the major gastrins in rat antrum, Component I, gastrin 34 ("big" gastrin), and gastrin 17-like peptides ("little" gastrin) was demonstrated after in vitro incubation of antral mucosal slices. The antral gastrin synthesis was stimulated by fundectomy, which ablates gastric acid secretion and thereby increases the gastrin concentration in plasma 20-fold. Tyrosine O-sulfation of gastrin was also demonstrated by incorporation of [35S]sulfate. Sulfate-labeled peptides precipitated by gastrin antisera coeluted with sulfated gastrin immunoreactivity on ion-exchange chromatography. Aryl-sulfatase treatment removed the [35S]sulfate from labeled gastrin and resulted in a change in elution pattern of the immunoreactivity to that of the unsulfated gastrins. The presence of 35S-labeled tyrosine O-sulfate residues was directly demonstrated by two-dimensional thin-layer electrophoresis after alkaline hydrolysis of [35S]sulfate-labeled gastrin. All the antral gastrins incorporated [35S]sulfate.
在大鼠胃窦黏膜切片进行体外孵育后,证实了标记氨基酸在大鼠胃窦主要胃泌素中的生物合成掺入,成分I,胃泌素34(“大”胃泌素)和胃泌素17样肽(“小”胃泌素)。胃底切除术刺激胃窦胃泌素合成,该手术消除胃酸分泌,从而使血浆中胃泌素浓度增加20倍。通过掺入[35S]硫酸盐也证实了胃泌素的酪氨酸O-硫酸化。胃泌素抗血清沉淀的硫酸盐标记肽在离子交换色谱上与硫酸化胃泌素免疫反应性共洗脱。芳基硫酸酯酶处理从标记的胃泌素中去除了[35S]硫酸盐,并导致免疫反应性的洗脱模式变为未硫酸化胃泌素的洗脱模式。在[35S]硫酸盐标记的胃泌素碱性水解后,通过二维薄层电泳直接证实了35S标记的酪氨酸O-硫酸酯残基的存在。所有胃窦胃泌素都掺入了[35S]硫酸盐。