%A Neeraj Dhaunta
%A Uzma Fatima
%A Purnananda Guptasarma
%O Copyright of this article belongs to Elsevier Science
%J Analytical biochemistry
%T N-Terminal sequencing by mass spectrometry through specific fluorescamine labeling of ?-amino groups before tryptic digestion.
%X 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 ?- and ?-amino groups associated with amino acids, in fact shows a specific reaction only with ?-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.
%N 2
%K  Fluorescamine adduction; Mass spectrometry; Protein sequencing; N-terminal sequencing; pH effects; ?-Amino group reactivity


%P 263-8
%V 408
%D 2011
%I Elsevier Science
%L open495