Selective N-terminal fluorescent labeling of proteins using 4-chloro-7-nitrobenzofurazan: A method to distinguish protein N-terminal acetylation

Lina F. Bernal-Perez, Laszlo Prokai, Youngha Ryu

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

Abstract

A fluorogenic derivatization method was developed to distinguish the protein N-terminal acetylation status. The unacetylated protein selectively reacted with 4-chloro-7-nitrobenzofurazan (NBD-Cl) at neutral pH to provide high fluorescence. In contrast, the protein with N-terminal acetylation was essentially nonfluorescent under the same conditions despite the presence of many internal lysine residues. Fluorescence of the NBD-labeled protein was very stable, and only micromolar concentrations of proteins were required for reliable detection. This method also provides a general and practical way to quantify proteins when their N-terminal amino group is available.

Original languageEnglish
Pages (from-to)13-15
Number of pages3
JournalAnalytical Biochemistry
Volume428
Issue number1
DOIs
StatePublished - 1 Sep 2012

Keywords

  • Fluorescent labeling
  • NBD-Cl
  • Protein N-terminal acetylation
  • Thymosin
  • Z-domain

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