Purification of porcine pancreatic kallikrein using various affinity chromatographies and intestinal absorption of the purified kallikrein.
For the simple purification of porcine pancreas kallikrein, various affinity chromatographies using L-prolyl-L-phenylalanyl-L-arginine (Pro-Phe-Arg-OH), tosyl-L-arginine (Tos-Arg-OH) and acetyl-L-phenylalanyl-L-arginine (Ac-Phe-Arg-OH) as ligands were examined. The purification was performed as follows using the extract from acetone powder of porcine pancreas as starting material; ammonium sulfate fractionation (30-80%), affinity chromatography (repeated) and gel filtration on Sephadex G-75, respectively. With these purification techniques, porcine pancreas kallikrein was purified about 2000-fold, respectively. From the results, these affinity chromatographies were considered to be effective for the purification of porcine pancreas kallikrein. Intestinal absorption of the purified kallikrein was also investigated using a rabbit. After the administration of the purified kallikrein into the intestinal tract, peripheral blood was taken from the ear vein and the plasma was assayed for Pro-Phe-Arg-7-amino-4-methylcoumarinamide hydrolytic activity. The concentration of the kallikrein in the blood was maximal at 3 hr after the administration and the absorption at this time amounted to about 0.07%.
D000818:Animals; D007611:Aprotinin / Q000494:pharmacology; D002846:Chromatography, Affinity; D007408:Intestinal Absorption; D007610:Kallikreins / Q000008:administration & dosage / Q000302:isolation & purification* / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology; D010179:Pancreas / Q000201:enzymology*; D011817:Rabbits; D013379:Substrate Specificity; D013552:Swine
D009842:Oligopeptides; C025273:prolyl-phenylalanyl-arginine-4-methylcoumaryl-7-amide; D007611:Aprotinin; D007610:Kallikreins