Booth A G, Kenny A J
Biochem J. 1976 Nov;159(2):395-407. doi: 10.1042/bj1590395.
The proteins of microvilli prepared from pig kidney were analysed by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. The typical pattern stained for protein revealed five major bands, four of which also stained for carbohydrate, and about 15 minor bands. For descriptive purposes the bands were designated numerically by their apparent molecular weights (X10(-3). Well-characterized proteins were identified with four of the five major bands. Dipeptidyl peptidase IV, a serine proteinase that may be specifically labelled with di-isopropyl [32P]phosphorofluoridate, was assigned to band 130. Aminopeptidase M was assigned to band 160, though when released from the membrane by a proteinase, this protein comprises three polypeptides each of lower apparent molecular weight than the native enzyme. Neutral endopeptidase can be assigned to band 95 and actin to band 42. The fifth major band (180) is an extrinsic glycoprotein that has not been identified with any microvillus enzyme activity. These four proteins contribute 21% of the microvillus-membrane protein. Kidney microvillus actin was characterized by a variety of properties and was similar to muscle actin. A computer analysis of the gel pattern indicates that it comprises 9.0% of the microvillus protein. Myosin is not present in the microvillus, but another protein associated with band 95, with properties that distinguish it from neutral endopeptidase, was tentatively identified as alpha-actinin. Alkaline phosphatase was identified as a monomeric polypeptide with an apparent molecular weight of 80000; it is a minor protein of the microvillus and is not discernible as a discrete band in the gel pattern. These and other results permit a model of the organization of the microvillus protein to be suggested. The computer program used has been deposited as Supplementary Publication SUP 50070 (12 pages) at the British Library Lending Division, Boston Spa. Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms given in Biochem. J. (1976) 153,5.
采用十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳法，对从猪肾制备的微绒毛蛋白进行了分析。蛋白质染色显示的典型图谱有五条主要条带，其中四条也被碳水化合物染色，还有约15条次要条带。为便于描述，这些条带按其表观分子量（×10⁻³）进行编号。已鉴定出五条主要条带中的四条具有特征明确的蛋白质。二肽基肽酶IV是一种丝氨酸蛋白酶，可用二异丙基[³²P]磷酰氟特异性标记，位于130条带。氨肽酶M位于160条带，不过当用蛋白酶从膜上释放时，该蛋白由三条表观分子量均低于天然酶的多肽组成。中性内肽酶位于95条带，肌动蛋白位于42条带。第五条主要条带（180）是一种外在糖蛋白，尚未鉴定出其具有任何微绒毛酶活性。这四种蛋白质占微绒毛膜蛋白的21%。肾微绒毛肌动蛋白具有多种特性，与肌肉肌动蛋白相似。对凝胶图谱的计算机分析表明，它占微绒毛蛋白的9.0%。微绒毛中不存在肌球蛋白，但与95条带相关的另一种蛋白质，其特性与中性内肽酶不同，暂被鉴定为α - 辅肌动蛋白。碱性磷酸酶被鉴定为一种表观分子量为80000的单体多肽；它是微绒毛中的一种次要蛋白质，在凝胶图谱中无法作为一条离散的条带分辨出来。这些及其他结果提示了一个微绒毛蛋白组织的模型。所使用的计算机程序已作为补充出版物SUP 50070（12页）存放在英国西约克郡韦瑟比波士顿温泉市大英图书馆出借部，邮编LS23 7BQ，可按《生物化学杂志》（1976年）第153卷第5期所规定的条件从该处获取复印件。
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