Synthesis of heptapeptides and analysis of sequence by tandem ion trap mass spectrometry |
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Authors: | Chenxi Jia Wei Qi Zhimin He Haoming Yang Bin Qiao |
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Institution: | (1) Chemical Engineering Research Center, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, People’s Republic of China |
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Abstract: | Two heptapeptides have been prepared by Fmoc methodology using Wang resin as solid support. For attachment of the first amino
acid, several coupling systems were evaluated, and DIC/DMAP system could give yields of >99% and low levels of racemization.
The selection of scavenger combination to deprotect side chains revealed that H2O/p-cresol was good at scavenging trityl and 1,2-ethanedithiol was highly efficient for scavenging t-butyl. Through shortening
the preactivation time to 5 min, the racemization which occurred during formation of amide bonds coupled by HBTU was minimized.
The crude peptides were characterized by RP-HPLC and MS, and sequenced by MS/MS to acquire reliable amino acid sequence information. |
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Keywords: | Peptide synthesis mass spectrometry peptide sequencing racemization RP-HPLC scavenger |
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