Enhanced a1 fragmentation for dimethylated proteins and its applications for N-terminal identification and comparative protein quantitation

Jue Liang Hsu, Shu-Hui Chen, Ding Tzai Li, Fong Ku Shi

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

In this work, dimethyl labeling at the protein level was developed to assist the fragmentation of intact proteins using the Q-TOF instrument. It was shown that a! ions were favorably enhanced upon collision-induced dissociation for dimethylated proteins with molecular mass below 20 kDa and without N-terminal modifications. This method is helpful in confirming proteolytic sites located at the N-terminus of proteins. Moreover, this labeling could be incorporated with stable isotopes for comparative profiling at the protein level, in which the heavily labeled and lightly labeled a1 ions were generated from the corresponding proteins upon high-voltage collisions in a broad mass region that covered all of the charge states of the proteins. Using hemoglobin as an example, a linear dynamic range from 1:1 to 1:20 was satisfactorily obtained with an R2 value greater than 0.99. This approach appears to be promising for top-down proteomics.

Original languageEnglish
Pages (from-to)2376-2383
Number of pages8
JournalJournal of Proteome Research
Volume6
Issue number6
DOIs
Publication statusPublished - 2007 Jun 1

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Proteins
Labeling
Ions
Molecular mass
Isotopes
Proteomics
Hemoglobins
Electric potential

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Chemistry(all)

Cite this

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abstract = "In this work, dimethyl labeling at the protein level was developed to assist the fragmentation of intact proteins using the Q-TOF instrument. It was shown that a! ions were favorably enhanced upon collision-induced dissociation for dimethylated proteins with molecular mass below 20 kDa and without N-terminal modifications. This method is helpful in confirming proteolytic sites located at the N-terminus of proteins. Moreover, this labeling could be incorporated with stable isotopes for comparative profiling at the protein level, in which the heavily labeled and lightly labeled a1 ions were generated from the corresponding proteins upon high-voltage collisions in a broad mass region that covered all of the charge states of the proteins. Using hemoglobin as an example, a linear dynamic range from 1:1 to 1:20 was satisfactorily obtained with an R2 value greater than 0.99. This approach appears to be promising for top-down proteomics.",
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Enhanced a1 fragmentation for dimethylated proteins and its applications for N-terminal identification and comparative protein quantitation. / Hsu, Jue Liang; Chen, Shu-Hui; Li, Ding Tzai; Shi, Fong Ku.

In: Journal of Proteome Research, Vol. 6, No. 6, 01.06.2007, p. 2376-2383.

Research output: Contribution to journalArticle

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AU - Shi, Fong Ku

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