Kinetic characterization of sequencing grade modified trypsin
- 2 June 2005
- journal article
- research article
- Published by Wiley in Proteomics
- Vol. 5 (9) , 2319-2321
- https://doi.org/10.1002/pmic.200401268
Abstract
Prior to analysis by mass spectrometry, protein samples are often digested. Maximizing the peptide yield from digestion can increase the number of peptides detected and the confidence in protein identification. To determine the optimal conditions for digestion, the Michaelis-Menten kinetic parameters for Promega sequencing grade modified trypsin were measured over a range of temperatures and pHs. The results indicate that an increase in digestion temperature above 37°C, the temperature traditionally used in digestion methods, could offer an increase in peptides detected.Keywords
This publication has 11 references indexed in Scilit:
- The preparation and properties of two new chromogenic substrates of trypsinPublished by Elsevier ,2004
- Optimized sample-processing time and peptide recovery for the mass spectrometric analysis of protein digestsJournal of the American Society for Mass Spectrometry, 2004
- Comparison of automated in‐gel digest methods for femtomole level samplesElectrophoresis, 2003
- Fast-Response Proteomics by Accelerated In-Gel Digestion of ProteinsAnalytical Chemistry, 2003
- Improvement of in‐gel digestion protocol for peptide mass fingerprinting by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometryRapid Communications in Mass Spectrometry, 2001
- Chemical modification of enzymes for enhanced functionalityCurrent Opinion in Biotechnology, 1999
- Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide GelsAnalytical Chemistry, 1996
- A continuous fluorescent assay for measuring protease activity using natural protein substrateAnalytical Biochemistry, 1991
- Fluorescein isothiocyanate-labeled casein assay for proteolytic enzymesAnalytical Biochemistry, 1984
- Stabilization of bovine trypsin by reductive methylationBiochimica et Biophysica Acta (BBA) - Protein Structure, 1977