Selective N-terminal fluorescent labeling of proteins using 4-chloro-7-nitrobenzofurazan: a method to distinguish protein N-terminal acetylation.
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.
Department of Chemistry, Texas Christian University, Fort Worth, TX 76129, USA.
Lina F
LF
Bernal-Perez
Laszlo
L
Prokai
Youngha
Y
Ryu
D009327:4-Chloro-7-nitrobenzofurazan / Q000737:chemistry / Q000378:metabolism*; D000107:Acetylation; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*
D005456:Fluorescent Dyes; D011506:Proteins; D009327:4-Chloro-7-nitrobenzofurazan