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Specifically, the system consisted of a Thermo Q-Exactive in line with an electrospray source and an Ultimate3000 series HPLC consisting of a binary pump, degasser and auto-sampler outfitted with an Xbridge Amide column (Waters) (dimensions: 3.0 100 mm and a 3.5-m particle size)
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Even though Tyr9 is not a base causing the unusual acidity of the thiol, it contributes to the mechanism as indicated by the low isomerase activity of the Tyr9Phe mutant, being only 2% of the wildtype activity (Pettersson and Mannervik, 2001)
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