Infrared-assisted proteolysis using trypsin-immobilized silica microspheres for peptide mapping.

Abstract:

:A novel proteolysis approach was developed by using infrared (IR) radiation and trypsin-immobilized silica microspheres. Protein solutions containing trypsin-immobilized microspheres in sealed transparent Eppendorf tubes were allowed to digest under an IR lamp at 37 degrees C. The feasibility and performance of the present proteolysis approach were demonstrated by the digestion of BSA and cytochrome c (Cyt-c) and the digestion time was significantly reduced to 5 min. The obtained digests were identified by MALDI-TOF-MS with the sequence coverages of 54% (BSA) and 83% (Cyt-c) that were better than those obtained by conventional in-solution tryptic digestion. The suitability of the new digestion approach to complex proteins was demonstrated by digesting human serum. The present proteolysis strategy is simple and efficient and will find a wide range of application in protein identification.

journal_name

Proteomics

journal_title

Proteomics

authors

Bao H,Lui T,Zhang L,Chen G

doi

10.1002/pmic.200800659

subject

Has Abstract

pub_date

2009-02-01 00:00:00

pages

1114-7

issue

4

eissn

1615-9853

issn

1615-9861

journal_volume

9

pub_type

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