Two-Dimensional Electrophoresis and Automated Microsequencing

Abstract

In organdies, cells and tissues of all organisms complex functions and metabolic reactions are to be maintained by proteins within and at the surface of each compartment. The number of proteins occuring in a biological compartment corresponds to the complexity of the functions it has to fulfill. Ribosomes as relatively high specialized compartments contain in the case of E.coli 54 different proteins, whereas estimations for the number of expressed genes in a typical human cell are in the range of about 5000. The human genome contains about 50000–100000 genes. For the registration and characterization of the components of these complex systems high resolution methods are necessary. The idea to characterize all components of a biological compartment became reality for the ribosomes when methods became available for the resolution of about 50 different proteins. These methods were first two-dimensional electrophoresis (2-DE)¹ and later also HPLC². Today, improved techniques of 2-DE play an important role to investigate the functional part of the genes, the proteins, within the human genome project. Furthermore changes in protein composition may be elucidated by subtractive analyses in different biological situations for example diseases or during differentiation.

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