Enhanced a1 Fragmentation for Dimethylated Proteins and Its Applications for N-Terminal Identification and Comparative Protein Quantitation
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- 作者：Jue-Liang Hsu ; Shu-Hui Chen ; Ding-Tzai Li ; Fong-Ku Shi
- 刊名：Journal of Proteome Research
- 出版年：2007
- 出版时间：June 2007
- 年：2007
- 卷：6
- 期：6
- 页码：2376 - 2383
- 全文大小：208K
- 年卷期：v.6,no.6(June 2007)
- ISSN：1535-3907
文摘
In this work, dimethyl labeling at the protein levelwas developed to assist the fragmentation of intactproteins using the Q-TOF instrument. It was shown thata1 ions were favorably enhanced upon collision-induceddissociation for dimethylated proteins with molecularmass below 20 kDa and without N-terminal modifications.This method is helpful in confirming proteolytic siteslocated at the N-terminus of proteins. Moreover, thislabeling could be incorporated with stable isotopes forcomparative profiling at the protein level, in which theheavily labeled and lightly labeled a1 ions were generatedfrom the corresponding proteins upon high-voltage collisions in a broad mass region that covered all of thecharge states of the proteins. Using hemoglobin as anexample, a linear dynamic range from 1:1 to 1:20 wassatisfactorily obtained with an R2 value greater than 0.99.This approach appears to be promising for top-downproteomics.