Dansyl-peptides matrix-assisted laser desorption/ionization mass spectrometric (MALDI-MS) and tandem mass spectrometric (MS/MS) features improve the liquid chromatography/MALDI-MS/MS analysis of the proteome.
Peptide tagging is a useful tool to improve matrix-assisted laser desorption/ionization tandem mass spectrometric (MALDI-MS/MS) analysis. We present a new application of the use of the dansyl chloride (DNS-Cl). DNS-Cl is a specific primary amine reagent widely used in protein biochemistry. It adds a fluorescent dimethylaminonaphthalene moiety to the molecule. The evaluation of MALDI-MS and MS/MS analyses of dansylated peptides shows that dansylation raises the ionization efficiency of the most hydrophilic species compared with the most hydrophobic ones. Consequently, higher Mascot scores and protein sequence coverage are obtained by combining MS and MS/MS data of native and tagged samples. The N-terminal DNS-Cl sulfonation improves the peptide fragmentation and promotes the generation of b-fragments allowing better peptide sequencing. In addition, we set up a labeling protocol based on the microwave chemistry. Peptide dansylation proved to be a rapid and cheap method to improve the performance of liquid chromatography (LC)/MALDI-MS/MS analysis at the proteomic scale in terms of peptide detection and sequence coverage.
Chiappetta
Giovanni
G
USR 3149 CNRS/ESPCI ParisTech, Ecole Supérieure de Physique et de Chimie Industrielles de Paris, Paris, France. giovanni.chiappetta@unina.it
Ndiaye
Sega
S
Demey
Emmanuelle
E
Haddad
Iman
I
Marino
Gennaro
G
Amoresano
Angela
A
Vinh
Joëlle
J
D000595:Amino Acid Sequence; D000818:Animals; D001920:Bradykinin / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D003619:Dansyl Compounds / Q000737:chemistry*; D004926:Escherichia coli / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D010449:Peptide Mapping / Q000379:methods; D020543:Proteome / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
D003619:Dansyl Compounds; D010446:Peptide Fragments; D020543:Proteome; D012710:Serum Albumin, Bovine; D014357:Trypsin; D001920:Bradykinin