4.5 Article Proceedings Paper

Kinetic characterization of sequencing grade modified trypsin

Journal

PROTEOMICS
Volume 5, Issue 9, Pages 2319-2321

Publisher

WILEY
DOI: 10.1002/pmic.200401268

Keywords

enzyme characterization; protein digestion; sample preparation; trypsin

Funding

  1. NIMH NIH HHS [R01MH59926] Funding Source: Medline

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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 degrees C, the temperature traditionally used in digestion methods, could offer an increase in peptides detected.

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