Latter G I, Burbeck S, Fleming J, Leavitt J
Clin Chem. 1984 Dec;30(12 Pt 1):1925-32.
We present a method that can, in principle, provide tentative identification of a substantial proportion of the polypeptides resolvable on a given two-dimensional electrophoresis gel. Computerized microdensitometry of autoradiograms from 20 gels labeled in turn with each of the 20 common amino acids provides the data for simultaneously measuring the amino acid composition of all polypeptides of interest on the gel. These compositions are then compared with computer data bases of known protein compositions. Similarity between a known and an unknown polypeptide with comparable molecular mass indicates a potential identification, which can then be confirmed with conventional techniques. We illustrate this technique by applying it to the identification of proteins in a transformed human cell line (HuT-14).
我们提出了一种方法,原则上能够对在给定二维电泳凝胶上可分辨的大部分多肽进行初步鉴定。用20种常见氨基酸依次对20块凝胶进行放射自显影片的计算机微密度测定,可提供数据用于同时测量凝胶上所有感兴趣多肽的氨基酸组成。然后将这些组成与已知蛋白质组成的计算机数据库进行比较。已知多肽与未知多肽之间具有可比分子量的相似性表明可能存在鉴定结果,随后可用传统技术进行确认。我们通过将该技术应用于转化人细胞系(HuT-14)中的蛋白质鉴定来阐述这一技术。