RESUMEN
Peptides obtained as a result of cyanogen bromide cleavage of the G-factor have been studied. All 12 peptides embracing the whole structure of fragment T4 have been isolated. For their amino acid sequence determination, cyanogen bromide peptides have been further cleaved with trypsin, chymotrypsin, thermolysin, staphylococcal glutamic protease and BNPS-skatole. The complete primary structure of 9 from 12 cyanogen bromide peptides has been determined.
Asunto(s)
Bromuro de Cianógeno , Escherichia coli/genética , Factores de Elongación de Péptidos/análisis , Secuencia de Aminoácidos , Cromatografía por Intercambio Iónico , Hidrólisis , Factor G de Elongación Peptídica , TripsinaRESUMEN
For fragment T4, obtained on limited trypsinolysis of the G-factor, the amino acid sequence embracing 76% of its structure has been determined by analysis of peptides resulting from the fragment T4 cleavage with staphylococcal glutamic protease. These data permitted to assemble into one polypeptide chain 7 out of 12 earlier characterized cyanogen bromide peptides contained in the fragment T4.
Asunto(s)
Endopeptidasas/farmacología , Escherichia coli/genética , Factores de Elongación de Péptidos/análisis , Fragmentos de Péptidos/análisis , Secuencia de Aminoácidos , Cromatografía Líquida de Alta Presión , Cromatografía por Intercambio Iónico , Bromuro de Cianógeno , Hidrólisis , Factor G de Elongación PeptídicaRESUMEN
Products of tryptic hydrolysis of the maleic anhydride modified fragment Th3 from limited thermolytic hydrolysis of the G-factor have been studied. Some short peptides which result from the trypsin action on the native G-factor molecule and belong to the fragment T4 obtained on limited trypsinolysis of the G-factor have been separated and their structure has been studied. As a result amino acid sequence has been determined by tryptic peptides containing 322 amino acid residues of the fragment T4 which makes up about 94% of its polypeptide chain.
Asunto(s)
Escherichia coli/genética , Factores de Elongación de Péptidos/análisis , Fragmentos de Péptidos/análisis , Tripsina , Secuencia de Aminoácidos , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Hidrólisis , Factor G de Elongación PeptídicaRESUMEN
The amino acid sequence of cysteine- and cystine-containing peptides resulting from cleavage of the G-factor by cyanogen bromide has been determined. For structure analysis cyanogen bromide peptides were further degradated using trypsin, chymotrypsin, thermolysin, staphylococcal glutamic protease, or limited acid hydrolysis. The products of the G-factor cleavage at Asp-Pro bonds were also studied. The obtained data together with those published earlier permitted to establish the complete primary structure of the elongation factor G. The polypeptide chain consists of 701 amino acid residues and has molecular mass of 77321,46.
Asunto(s)
Bromuro de Cianógeno , Escherichia coli/genética , Factores de Elongación de Péptidos/análisis , Fragmentos de Péptidos/análisis , Secuencia de Aminoácidos , Cromatografía en Agarosa , Cromatografía en Gel , Cromatografía Líquida de Alta Presión , Hidrólisis , Factor G de Elongación Peptídica , Reactivos de SulfhidriloRESUMEN
We suggest a simple approach to localization of antigenic determinants for the monoclonal antibody 13B1 raised against recombinant human interleukin-2. The approach is based on the limited trypsin proteolysis, peptide separation by the O'Farrell method and identification of the peptides, interacting with monoclonal antibodies, and comparison of the charge and length of these peptides with the corresponding values of theoretically possible peptides.