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Lookup NU author(s): Dr Norah O'Farrell
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We have developed an electrospray ionisation mass spectrometry (ESI-MS) technique that can be applied to rapidly determine the number of intact active sites in proteins. The methodology relies on inhibiting the protein with an active-site irreversible inhibitor and then using ESI-MS to determine the extent of inhibition. We have applied this methodology to a test system: a serine protease, subtilisin Carlsberg, and monitored the extent of inhibition by phenylmethylsulfonyl fluoride (PMSF), an irreversible serine hydrolase inhibitor as a function of the changes in immobilisation and hydration conditions. Two types of enzyme preparation were investigated, lyophilised enzymes and protein-coated microcrystals (PCMC). © 2006 Wiley Periodicals, Inc.
Author(s): O'Farrell N, Kreiner M, Moore BD, Parker M-C
Publication type: Article
Publication status: Published
Journal: Biotechnology and Bioengineering
ISSN (print): 0006-3592
ISSN (electronic): 1097-0290
Publisher: John Wiley & Sons, Inc.
PubMed id: 16917861
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