Issue Date: July 21, 2008
Pressure Cooker Proteomics
Chopping collections of proteins into manageable peptide pieces is one of the most time-consuming parts of proteomics. But with the help of some pressure, that sluggish enzyme-catalyzed step, which normally takes four hours or longer, can be carried out in just one minute (J. Proteome Res., DOI: 10.1021/pr7008077). Richard D. Smith and colleagues at Pacific Northwest National Laboratory applied pressure pulses to a complex bacterial protein mixture in the presence of trypsin, a protein-clipping enzyme that's commonly used for proteome sample preparation. After one minute, they analyzed the peptide products by mass spectrometry and compared the results with those from a conventional hours-long protocol with trypsin. The collection of peptides obtained under pressure was comparable with results from the traditional method. The researchers' additional experiments indicate that increased pressure denatures proteins, giving trypsin room to work quickly. Microwave- and ultrasound-based techniques have already been used to speed up the trypsin step, but the pressure-based technique simplifies sample preparation, making it a promising candidate for high-throughput technology, the authors write.
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