Effect of the hydrolysis method on the determination of the amino acid composition of proteins

Journal of Chromatography. a
M WeissM Fountoulakis

Abstract

Fast and reproducible separation and determination of amino acids serves the economical and reliable characterization and quantification of peptides and proteins as well as the identification of proteins by amino acid composition analysis on a large-scale. A prerequisite of a successful compositional analysis is a complete hydrolysis of the peptides and proteins and a quantitative recovery of the residues in the hydrolyzate. We investigated the effect of different acid-hydrolysis methods on the compositional analysis of known proteins in solution and after blotting onto polyvinylidene difluoride membranes and worked out the conditions for the processing of large numbers of samples. The reliability of each method was studied by introducing the analysis data into the AACompIdent software and deducing the protein identification scores. All acid-hydrolysis methods delivered reliable analysis data. The most accurate data were provided by conventional, thermal hydrolysis of proteins in solution in the presence of methanesulfonic acid, closely followed by hydrolysis with hydrochloric acid and microwave radiation-dependent hydrolysis with hydrochloric or methanesulfonic acid, respectively. For blotted proteins, conventional hydrolysis ...Continue Reading

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Citations

May 24, 2011·Analytical and Bioanalytical Chemistry·Orsolya Egressy-MolnárMihály Dernovics
Mar 12, 2003·Journal of Hazardous Materials·E NeyensB De heyder
Oct 31, 2009·Journal of Pharmaceutical and Biomedical Analysis·Uwe ConradGerhard K E Scriba
Feb 20, 2007·Journal of Biochemical and Biophysical Methods·Shi-Zhong YangBo-Zhong Mu
Jul 20, 2014·Biologicals : Journal of the International Association of Biological Standardization·Weslen S Vedakumari, Thotapalli P Sastry
Sep 2, 2003·Journal of Pharmaceutical Sciences·Lei WanTse Wen Chang
Nov 7, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Tobias LangrockRalf Hoffmann
Jul 19, 2006·Journal of Pharmaceutical and Biomedical Analysis·Silvia De Boni, Gerhard K E Scriba
May 11, 2004·Mass Spectrometry Reviews·Michael Fountoulakis
May 18, 2018·Food Science and Technology International = Ciencia Y Tecnología De Los Alimentos Internacional·F KermicheA Romero
May 25, 2010·Journal of Dairy Science·R MarinoM Albenzio
Feb 16, 2000·Electrophoresis·H LangenM Fountoulakis
Nov 26, 2009·Current Protocols in Protein Science·Shane M Rutherfurd, G Sarwar Gilani
Jan 31, 2004·The Analyst·Mireia Fernandez OcañaRaj Patel
Mar 1, 2018·Comprehensive Reviews in Food Science and Food Safety·Olga Luisa TavanoRoberto Fernandez-Lafuente
Jan 23, 1999·Journal of Chromatography. a·M Fountoulakis, H W Lahm
Aug 20, 2003·Journal of Chromatography. a·Katrin SalchertCarsten Werner
Nov 23, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mallikarjun BadadaniK Taranath Shetty
Nov 14, 2014·Journal of Agricultural and Food Chemistry·Yong Guan ChuaLai Peng Leong

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