Systematic analysis of the total proteins of a mammalian organism: principles, problems and implications for sequencing the human genome.
High-resolution two-dimensional electrophoresis (2-DE) has reached a technological level that allows us to resolve most of the numerous unknown protein species of a mammalian organism if appropriate strategies are used. We will discuss the problems of classification and characterization of proteins and propose a systematic approach to the analysis of the total protein complex. Both a comprehensive as well as a pragmatic approach towards systematic analysis have been considered. A "complex protein database" is suggested and considered with regard to various uses. A systematic analysis of the mouse proteins has been started and some of the preliminary results are summarized here. In particular, genetic properties of the proteins were investigated and are presented in order to demonstrate the significance of a systematic analysis of proteins for research and practical application (e.g. mutagenicity testing). A concept is presented for sequencing the coding DNA of mouse and man, starting with a systematic analysis of mouse proteins and then using two recently developed methods - microsequencing of proteins from spots of 2-DE protein patterns, and utilization of the relatively short N-terminal sequences obtained - to produce the corresponding cDNA's of these proteins.
Klose
J
J
Institut für Humangenetik, Freie Universität Berlin, Federal Republic of Germany.
D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D004247:DNA / Q000032:analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007256:Information Systems; D051379:Mice; D011506:Proteins / Q000032:analysis* / Q000145:classification
D011506:Proteins; D004247:DNA