Kumazaki T, Kobayashi M, Ishii S
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Hokkaido University, Sapporo, Japan.
J Mol Recognit. 1988 Apr;1(2):93-8. doi: 10.1002/jmr.300010207.
Anhydroelastase was effectively isolated by a single operation of affinity chromatography from a complex mixture produced by phenylmethylsulfonylation and alkaline treatment of porcine pancreatic elastase. The adsorbent used for the chromatography was 6-aminohexanoyl-trialanine, which corresponds to a product of elastase action, immobilized on Sepharose 4B. Successful resolution by the operation indicated that this immobilized ligand possesses the highest affinity for anhydroelastase among various proteins including regenerated elastase in the mixture. Comparative affinity chromatography on immobilized anhydroelastase and on immobilized native elastase further confirmed the stronger interaction of anhydroelastase with the product-type peptides. Immobilized anhydroelastase was also found to be useful in the purification and search for naturally occurring proteinase inhibitors.
通过亲和色谱的单次操作，从猪胰弹性蛋白酶经苯甲基磺酰化和碱处理产生的复杂混合物中有效地分离出了脱水电弹性蛋白酶。用于色谱的吸附剂是固定在琼脂糖4B上的6-氨基己酰-三丙氨酸，它对应于弹性蛋白酶作用的产物。该操作的成功分离表明，在包括混合物中的再生弹性蛋白酶在内的各种蛋白质中，这种固定化配体对脱水电弹性蛋白酶具有最高的亲和力。在固定化脱水电弹性蛋白酶和固定化天然弹性蛋白酶上进行的比较亲和色谱进一步证实了脱水电弹性蛋白酶与产物型肽之间更强的相互作用。还发现固定化脱水电弹性蛋白酶可用于纯化和寻找天然存在的蛋白酶抑制剂。
