Identification of proteins by their amino acid composition: an evaluation of the method.

摘要: Expression of different genomes can be studied by high-resolution two-dimensional electrophoresis (2-D PAGE). To help these studies, reference maps biological tissues and fluids have been built found in the SWISS-2DPAGE database, accessible via World Wide Web network on ExPASy molecular biology server. Different techniques were used to identify polypeptides. At present time, method considered fastest most cost-effective is amino acid composition analysis (AAC). Proteins, transferred onto polyvinylidene (PVDF) membranes, submitted vapor-phase hydrolysis, derivatized with 9-fluorenylmethyl chloroformate (FMOC) separated an ODS-Hypersil column. Identification was obtained using program ‘AACompIdent’ available from ExPASy. In this work, experimental parameters, such as contamination, reproducibility accuracy, assessed. First, it has that a major source contamination human keratin. Next, acids classified into ‘reliable’ ‘nonreliable’. Accordingly, ‘bias’ ‘weights’ defined for each acid, which could set program. Finally, examples identification, including use Edman degradation sequence tagging, are described.