Dhaunta, Neeraj and Fatima, Uzma and Guptasarma, Purnananda (2011) N-Terminal sequencing by mass spectrometry through specific fluorescamine labeling of α-amino groups before tryptic digestion. Analytical biochemistry, 408 (2). pp. 263-8. ISSN 1096-0309
Full text not available from this repository. (Request a copy)Abstract
We present a single-step procedure for the specific mass labeling of unblocked protein N termini. We show that the dye fluorescamine, which is commonly assumed to require mildly alkaline conditions for undergoing a nonspecific reaction with α- and ε-amino groups associated with amino acids, in fact shows a specific reaction only with α-amino groups present at protein N termini when mildly acidic conditions are used. We use this finding to label, identify, and sequence the trypsinolysis-derived N-terminal peptide of lysozyme, using only mass spectrometry, to illustrate how this method could be used with other proteins.
| Item Type: | Article |
|---|---|
| Additional Information: | Copyright of this article belongs to Elsevier Science |
| Uncontrolled Keywords: | Fluorescamine adduction; Mass spectrometry; Protein sequencing; N-terminal sequencing; pH effects; α-Amino group reactivity |
| Subjects: | Q Science > QD Chemistry |
| Depositing User: | Dr. K.P.S.Sengar |
| Date Deposited: | 08 Dec 2011 18:57 |
| Last Modified: | 08 Dec 2011 18:57 |
| URI: | http://crdd.osdd.net/open/id/eprint/495 |
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