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Design and synthesis of a solid‐phase fluorescent mass tag
Author(s) -
Shi Yang,
Xiang Rong,
Horváth Csaba,
Wilkins James A.
Publication year - 2005
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200500115
Subject(s) - chemistry , cysteine , fluorescence , reagent , peptide , combinatorial chemistry , chromatography , solid phase synthesis , lysine , amino acid , mass spectrometry , biochemistry , organic chemistry , physics , quantum mechanics , enzyme
In this study, we demonstrate the design of a new solid‐phase fluorescent mass tag (FMT) that contains the following features: (1) the FMT is synthesized using Fmoc chemistry which is simple, rapid, and cost‐effective; (2) lysine is used as a uniformly labeled amino acid (using stable isotopes) to allow 8 Da difference between “heavy” and “light” tags; (3) a fluorescent molecule is coupled to the isotope tag that allows a tagged peptide to be detected by online fluorescence; and (4) an iodoacetyl reactive group provides cysteine reactivity. Using MALDI‐TOF MS and HPLC, we show that the FMT reagent can be used to label standard cysteine‐containing peptides as well as cysteine‐containing peptides from a BSA tryptic digest.