A new generation of cross-linkers for selective detection by MALDI MS.
We designed a new cross-linker bearing a CHCA moiety. The use of the CHCA-tagged cross-linker JMV 3378 in conjunction with a neutral MALDI matrix alpha-cyano-4-hydroxycinnamic methyl ester enabled specific signal enhancement in MALDI-TOF MS of cross-link containing peptides. Discrimination between modified and non-modified peptides can be achieved by comparison of two spectra, one using CHCA and the other using the alpha-cyano-4-hydroxycinnamic methyl ester matrix. The methodology was validated using cytochrome c and apo-myoglobine as model proteins.
Paramelle
David
D
Institut des Biomolécules Max Mousseron, UMR 5247 CNRS, Universités Montpellier 1 et 2, Montpellier, France.
Cantel
Sonia
S
Enjalbal
Christine
C
Amblard
Muriel
M
Forest
Eric
E
Heymann
Michaël
M
Geourjon
Christophe
C
Martinez
Jean
J
Subra
Gilles
G
D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000032:analysis; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D045304:Cytochromes c / Q000032:analysis; D006736:Horses; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009211:Myoglobin / Q000032:analysis; D010455:Peptides / Q000032:analysis; D011487:Protein Conformation; D011506:Proteins / Q000032:analysis*; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001059:Apoproteins; D003432:Cross-Linking Reagents; D009211:Myoglobin; D010455:Peptides; D011506:Proteins; C010857:apomyoglobin; D045304:Cytochromes c