Hänsler M, Appelt G, Rogos R
Forschungslabor Gastroenterologie, Medizinisch-Poliklinischen Instituts, Universität Leipzig.
Z Med Lab Diagn. 1991;32(3-4):173-9.
After isoelectric focusing of proteins of human pure pancreatic juice following digestive enzymes and zymogens were identified by substrate specific detection methods: in the anodic range trypsinogen 1, and acidic form of proelastase and procarboxypeptidase A; in the pH--range of 5.0-6.0 a trypsinogen--like protein and the secretory trypsin inhibitor; in the pH--range of 6.0-7.0 one form of each of procarboxypeptidase B, trypsinogen 2, lipase, chymotrypsinogen and two forms of alpha-amylase and in the cathodic range an activated form of procarboxypeptidase B, one form of prophospholipase A2 and an additional form of proelastase. In order to characterize glycoproteins the IEF separated pancreatic proteins were transferred onto nitrocellulose membranes. Using the periodic acid/Schiff's reagent (PAS) procedure five PAS-positive proteins (pl s 4.5; 6.0; 6.6; 6.9 and 7.3) were detected. Three Con A-binding proteins (pl s 6.0; 6.6 and 6.9) were identified by incubating the blot in Con A followed by peroxidase and o-phenylendiamin as a chromogenic peroxidase substrat.
通过底物特异性检测方法对人纯胰液中的蛋白质进行等电聚焦后，鉴定出消化酶和酶原：在阳极区域有胰蛋白酶原1、弹性蛋白酶原的酸性形式和羧肽酶原A；在pH值5.0 - 6.0范围内有类似胰蛋白酶原的蛋白质和分泌型胰蛋白酶抑制剂；在pH值6.0 - 7.0范围内有羧肽酶原B、胰蛋白酶原2、脂肪酶、糜蛋白酶原的各一种形式以及两种形式的α-淀粉酶，在阴极区域有羧肽酶原B的活化形式、磷脂酶原A2的一种形式和弹性蛋白酶原的另一种形式。为了鉴定糖蛋白，将等电聚焦分离的胰腺蛋白质转移到硝酸纤维素膜上。使用过碘酸/席夫试剂（PAS）程序检测到五种PAS阳性蛋白质（等电点分别为4.5、6.0、6.6、6.9和7.3）。通过将印迹与伴刀豆球蛋白A孵育，然后用过氧化物酶和邻苯二胺作为显色过氧化物酶底物，鉴定出三种伴刀豆球蛋白A结合蛋白（等电点分别为6.0、6.6和6.9）。
