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Modifications in Trypsin Digestion Protocol for Increasing the Efficiency and Coverage

Syal, Kirtimaan and Tadala, Raghu (2015) Modifications in Trypsin Digestion Protocol for Increasing the Efficiency and Coverage. In: PROTEIN AND PEPTIDE LETTERS, 22 (4). pp. 372-378.

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Abstract

Standard trypsin digestion protocol of proteins followed by MALDI-MS analysis has been realized as an important tool for the identification and characterization of proteins. In this article, we proposed the elimination of the step of `staining/de-staining of gel pieces' in in-gel digestion protocol in order to improve the efficiency of trypsin digestion. Coomassie dye is known to interfere with digestion of proteins by trypsin and the procedure of staining-de-staining could result in loss of photoaffinity probe, post translational modifications and catalytic activities of enzymes. Further, we studied parameters like hydrophobicity and isoelectric point, and attempted to quantitatively relate it to the efficiency of trypsin digestion. We suggest that properties of proteins should be considered and trypsin digestion protocol should be appropriately modified as per sequence and other information.

Item Type: Journal Article
Additional Information: Copy right for this article belongs to the BENTHAM SCIENCE PUBL LTD, EXECUTIVE STE Y-2, PO BOX 7917, SAIF ZONE, 1200 BR SHARJAH, U ARAB EMIRATES
Keywords: Bottom-up approach; hydrophobicity; isoelectric point; post-translational modifications; staining/de-staining; trypsin digestion
Department/Centre: Division of Biological Sciences > Molecular Biophysics Unit
Depositing User: Id for Latest eprints
Date Deposited: 06 May 2015 05:37
Last Modified: 06 May 2015 05:37
URI: http://eprints.iisc.ac.in/id/eprint/51502

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