Sulfated compounds in the zymogen granules of the guinea pig pancreas
Sulfated compounds in the zymogen granules of the guinea pig pancreas
复制标题
豚鼠胰腺酶原颗粒中的硫酸化化合物
作者：
H. Reggio;G. Palade
用离体培养的小小叶系统，在含~（35）SO_4 ~（-2）的KRB重碳酸盐培养基（KRB）中孵育60 min，通过光（LM）和电镜（EM）放射自显影研究了硫酸盐掺入豚鼠胰腺的情况。在腺泡细胞，检查EM放射自显影，标记被发现集中在高尔基体元素（包括冷凝空泡）和酶原颗粒。35 SO 4（-2）也被整个胰管系统的上皮细胞掺入，在主管上皮中的掺入量惊人地高。在所有导管上皮细胞中，高尔基复合体和质膜上均出现放射自显影颗粒。由于不能排除导管上皮对排出分泌物中发现的硫酸化化合物的贡献，因此使用纯化的酶原颗粒级分作为分离腺泡来源的硫酸化化合物的源材料。通过放射自显影术（切片颗粒）确定酶原颗粒和浓缩空泡中存在35 S-放射性。从该酶原颗粒级分的裂解物中，通过凝胶过滤在允许其与大量分泌蛋白质满意分离的条件下分离低等电点和高分子量的可溶性硫酸化化合物。
Sulfate incorporation into the guinea pig pancreas was investigated by light (LM) and electron microscope (EM) autoradiography using a system of minilobules incubated in vitro for 60 min in Krebs-Ringer bicarbonate medium (KRB) containing 35SO4(-2). In acinar cells, examined by EM autoradiography, the label was found concentrated over Golgi elements (including condensing vacuoles) and zymogen granules. 35SO4(-2) was also incorporated by the epithelial cells of the entire pancreatic duct system, the incorporation being surprisingly high in the epithelium of the major ducts. In all ductal epithelia, autoradiographic grains appeared over the Golgi complex and the plasmalemma. Since a contribution of duct epithelium to the sulfated compounds found in the discharged secretion could not be ruled out, a purified zymogen granule fraction was used as a source material for the isolation of sulfated compounds of acinar origin. The presence of 35S- radioactivity in the zymogen granules and condensing vacuoles of this fraction was ascertained by autoradiography (of sectioned pellets). From a lysate of this zymogen granule fraction, a soluble sulfated compound of low isoelectric point and high molecular weight was isolated by gel filtration under conditions that allowed its satisfactory separation from the bulk of the secretory proteins.