3-Aminoquinoline/p-coumaric acid as a MALDI matrix for glycopeptides, carbohydrates, and phosphopeptides.
Glycosylation and phosphorylation are important post-translational modifications in biological processes and biomarker research. The difficulty in analyzing these modifications is mainly their low abundance and dissociation of labile regions such as sialic acids or phosphate groups. One solution in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry is to improve matrices for glycopeptides, carbohydrates, and phosphopeptides by increasing the sensitivity and suppressing dissociation of the labile regions. Recently, a liquid matrix 3-aminoquinoline (3-AQ)/α-cyano-4-hydroxycinnamic acid (CHCA) (3-AQ/CHCA), introduced by Kolli et al. in 1996, has been reported to increase sensitivity for carbohydrates or phosphopeptides, but it has not been systematically evaluated for glycopeptides. In addition, 3-AQ/CHCA enhances the dissociation of labile regions. In contrast, a liquid matrix 1,1,3,3-tetramethylguanidium (TMG, G) salt of p-coumaric acid (CA) (G3CA) was reported to suppress dissociation of sulfate groups or sialic acids of carbohydrates. Here we introduce a liquid matrix 3-AQ/CA for glycopeptides, carbohydrates, and phosphopeptides. All of the analytes were detected as [M + H](+) or [M - H](-) with higher or comparable sensitivity using 3-AQ/CA compared with 3-AQ/CHCA or 2,5-dihydroxybenzoic acid (2,5-DHB). The sensitivity was increased 1- to 1000-fold using 3-AQ/CA. The dissociation of labile regions such as sialic acids or phosphate groups and the fragmentation of neutral carbohydrates were suppressed more using 3-AQ/CA than using 3-AQ/CHCA or 2,5-DHB. 3-AQ/CA was thus determined to be an effective MALDI matrix for high sensitivity and the suppression of dissociation of labile regions in glycosylation and phosphorylation analyses.
Koichi Tanaka Laboratory of Advanced Science and Technology, Shimadzu Corporation , 1, Nishinokyo-Kuwabaracho, Nakagyo-ku, Kyoto 604-8511, Japan.
Yuko
Y
Fukuyama
Natsumi
N
Funakoshi
Kohei
K
Takeyama
Yusaku
Y
Hioki
Takashi
T
Nishikaze
Kaoru
K
Kaneshiro
Shin-Ichirou
S
Kawabata
Shinichi
S
Iwamoto
Koichi
K
Tanaka
D000595:Amino Acid Sequence; D000818:Animals; D002241:Carbohydrates / Q000032:analysis* / Q000235:genetics; D002417:Cattle; D003373:Coumaric Acids / Q000737:chemistry*; D006020:Glycopeptides / Q000032:analysis* / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010748:Phosphopeptides / Q000032:analysis*; D011422:Propionates; D015363:Quinolones / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
C560164:3-aminoquinolein-2-one; D002241:Carbohydrates; D003373:Coumaric Acids; D006020:Glycopeptides; D010748:Phosphopeptides; D011422:Propionates; D015363:Quinolones; C495469:p-coumaric acid