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Direct,simple derivatization of disulfide bonds in proteins with organic mercury in alkaline medium without any chemical pre-reducing agents |
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| Authors: | Beatrice Campanella Massimo Onor Carlo Ferrari Alessandro D’Ulivo Emilia Bramanti |
| Institution: | 1. National Research Council of Italy, C.N.R., Istituto di Chimica dei Composti Organo Metallici-ICCOM- UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa, Italy;2. National Research Council of Italy, C.N.R., Istituto Nazionale di Ottica, INO-UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa, Italy |
| Abstract: | In this work we have studied the derivatization of protein disulfide bonds with p-Hydroxymercurybenzoate (pHMB) in strong alkaline medium without any preliminary reduction. The reaction has been followed by the determination of the protein–pHMB complex using size exclusion chromatography coupled to a microwave/UV mercury oxidation system for the on-line oxidation of free and protein-complexed pHMB and atomic fluorescence spectrometry (SEC–CVG–AFS) detection. The reaction has been optimized by an experimental design using lysozyme as a model protein and applied to several thiolic proteins. |
| Keywords: | p-Hydroxymercuribenzoic acid Proteins Disulfide bond derivatization Alkaline medium Atomic fluorescence spectrometry |
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