Tweeten K A, Bulla L A, Consigli R A
J Virol. 1978 Jun;26(3):703-11.
An alkaline protease was found to be associated with the granulosis virus of the Indian meal moth. Plodia interpunctella. The protease was located within the protein matrix of the occluded virus and hydrolyzed the major constituent of this matrix, a 28,000-dalton protein (granulin), to a mixture of polypeptides ranging in molecular weight from 10,000 to 27,000. A rapid, sensitive assay for the protease was developed using radioactively labeled granulosis virus as substrate. With this assay, the proteolytic activity could be detected by measuring the release of acid-soluble peptides from the labeled virus. The protease had a pH optimum of 10.5 and a temperature optimum of 40 degrees C and was inhibited by diisopropyl phosphorofluoridate, phenylmethylsulfonyl fluoride, and L-(1-tosylamido-2-phenyl) ethyl chloromethyl ketone. Purification of the protease from matrix protein was achieved by anion-exchange and gel permeation chromatography. The molecular weight of the isolated protease, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration, was approximately 14,000.
发现一种碱性蛋白酶与印度谷螟（Plodia interpunctella）的颗粒体病毒有关。该蛋白酶位于包涵体病毒的蛋白质基质中，可将这种基质的主要成分——一种28,000道尔顿的蛋白质（颗粒体蛋白）水解为分子量在10,000至27,000之间的多肽混合物。利用放射性标记的颗粒体病毒作为底物，开发了一种快速、灵敏的蛋白酶检测方法。通过这种检测方法，可通过测量标记病毒中酸溶性肽的释放来检测蛋白水解活性。该蛋白酶的最适pH为10.5，最适温度为40℃，并受到二异丙基氟磷酸酯、苯甲基磺酰氟和L-（1-甲苯磺酰氨基-2-苯基）乙基氯甲基酮的抑制。通过阴离子交换和凝胶渗透色谱法从基质蛋白中纯化了该蛋白酶。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和凝胶过滤测定，分离出的蛋白酶分子量约为14,000。
