N-Terminal sequencing by mass spectrometry through specific fluorescamine labeling of α-amino groups before tryptic digestion.
IR@IMTECH: CSIR-Institute of Microbial Technology, Chandigarh
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| Title |
N-Terminal sequencing by mass spectrometry through specific fluorescamine labeling of α-amino groups before tryptic digestion.
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| Creator |
Dhaunta, Neeraj
Fatima, Uzma Guptasarma, Purnananda |
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| Subject |
QD Chemistry
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| Description |
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.
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| Publisher |
Elsevier Science
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| Date |
2011-01-15
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| Type |
Article
PeerReviewed |
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| Relation |
http://www.sciencedirect.com/science/article/pii/S0003269710005853
http://crdd.osdd.net/open/495/ |
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| Identifier |
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
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