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Infrared‐assisted proteolysis using trypsin‐immobilized silica microspheres for peptide mapping

2009; Wiley; Volume: 9; Issue: 4 Linguagem: Inglês

10.1002/pmic.200800659

1615-9861

Huimin Bao, Ting Lui, Luyan Zhang, Gang Chen,

Advanced Proteomics Techniques and Applications

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°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.