Rapid and Sensitive Amino-Acid Sequencing of Cloning Thermus thermophilus HB8 Ferredoxin by Proteomics

Kaneko, Maki; Masui, Ryoji; Ake, Kojiro; Kousumi, Yukihide; Kuramitsu, Seiki; Yamaguchi, Minoru; Kuyama, Hiroki; Ando, Eiji; Norioka, Shigemi; Nakazawa, Takashi; Okamura, Taka‐aki; Yamamoto, H.; Ueyama, Norikazu

doi:10.1021/pr049929z

Cited by 7 publications

(4 citation statements)

References 15 publications

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“…The peak pattern and separation can be adjusted freely by a combination of charge tags labeled with heavy and light isotopes . Some metal complexes satisfy the criteria of rendering peptides with charge and characteristic peak pattern; a ruthenium complex has been introduced to the N-terminus of a large protein after being guanidinated at the ε-amino groups (Kaneko et al, 2004).…”

Section: N-terminal Modification For Tandem Msmentioning

confidence: 99%

Chemistry in Proteomics: An Interplay between Classical Methods in Chemical Modification of Proteins and Mass Spectrometry at the Cutting Edge

Nakazawa

2006

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This review deals with chemical approaches that facilitate proteomics research. The methods include chemical cleavage of peptide bonds, stepwise degradation of proteins, and site-specific modification enabling the discrimination between functional groups. These reactions could enhance the efficacy of mass spectrometry (MS) for identification, quantification, and sequencing of peptides and proteins. Particular stress is laid on the effect of each modification on mass spectra mainly with respect to the enhancement of peak intensities, the clarity of spectra, and the suppression of ambiguity in peak assignment. Chemical modifications targeting the terminal free amino and carboxyl groups are reviewed, because the charge or isotopic tagging on N-and C-termini has proved effective to identify a protein based on the terminal amino acid sequencing with tandem MS in combination with database searching. Such a tag that is designed to provide a peak with a characteristic sensitivity or pattern could corroborate the implication of mass spectra and facilitate the de novo protein sequencing of mature proteins, which are often subjected to post-translational modifications.

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Section: N-terminal Modification For Tandem Msmentioning

confidence: 99%

Chemistry in Proteomics: An Interplay between Classical Methods in Chemical Modification of Proteins and Mass Spectrometry at the Cutting Edge

Nakazawa

2006

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“…This reagent has a characteristic isotope distribution pattern (Fig. 1, bottom) and yields intense peaks in MS analysis 21–23. Intense <Ru>‐CO‐labeled N‐terminal fragment peaks were exclusively detected by matrix‐assisted laser desorption/ionization post‐source decay (MALDI‐PSD).…”

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Simultaneous detection of N‐terminal fragment ions in a protein mixture using a ruthenium(II) complex

Ito

Okamura

Yamamoto

et al. 2007

Rapid Comm Mass Spectrometry

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Use of a bis(terpyridine)ruthenium(II) derivative as an N-terminal labeling reagent resulted in the simultaneous detection and individual determination of all the N-terminal fragments of the proteins in a mixture without requiring any separation. All of the N-termini of the guanidinated proteins were labeled selectively by the ruthenium complex (-CO-labeling). After chymotryptic digestion, the fragments were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and post-source decay (PSD). The -CO moiety exclusively enhanced N-terminal fragment ions in mass spectra and enabled easy N-terminal sequencing. In a mixture containing three different proteins (lysozyme, ubiquitin, and insulin), all of the N-terminal fragment ions labeled with the ruthenium complex were found to produce uniformly intense peaks without the detection of the other unlabeled fragments. The N-terminal sequences of these ions were determined individually by PSD analysis. Application to unknown proteins from Thermus thermophilus HB8 with two-dimensional electrophoretic separation resulted in the successful determination of the N-terminal sequence and easy identification of the target protein.

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“…37,38 The reagent has a characteristic isotope pattern and yields highly intense peaks in MS analysis. [37][38][39][40][41] By using this reagent for amino acid sequencing of peptides or proteins, we selectively detected N-terminal fragment ions with no C-terminal fragment ion. N-Terminal labeling with the reagent 〈Ru〉-CO achieves rapid determination of the Nterminal amino acid sequence.…”

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“…Recently, we demonstrated protein sequencing using an active ester of bis(terpyridine)ruthenium(II) derivative (Figure ) as a novel N-terminal labeling reagent. , The reagent has a characteristic isotope pattern and yields highly intense peaks in MS analysis. − By using this reagent for amino acid sequencing of peptides or proteins, we selectively detected N-terminal fragment ions with no C-terminal fragment ion. N-Terminal labeling with the reagent 〈Ru 〉 -CO achieves rapid determination of the N-terminal amino acid sequence .…”

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Distinction of Leu and Ile Using a Ruthenium(II) Complex by MALDI-LIFT-TOF/TOF-MS Analysis

et al. 2005

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The novel N-terminal labeling method using a ruthenium(II) complex derivative characteristically indicated a(n) and d(n) (N-terminal) fragment ions in high sensitivity by MS/MS analysis (MALDI-LIFT or ESI-CID). Although these fragment ions depended on a fragmentation process by MS/MS analytical methods to some degree, each case indicated similar side-chain cleavage patterns. The labeling method allows accurate distinction of amino acid residues by MS/MS analysis even if the residues are structural isomers such as leucine and isoleucine. The method was applied to long-chain peptides and provided easy and rapid N-terminal sequencing.

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Rapid and Sensitive Amino-Acid Sequencing of Cloning Thermus thermophilus HB8 Ferredoxin by Proteomics

Cited by 7 publications

References 15 publications

Chemistry in Proteomics: An Interplay between Classical Methods in Chemical Modification of Proteins and Mass Spectrometry at the Cutting Edge

Chemistry in Proteomics: An Interplay between Classical Methods in Chemical Modification of Proteins and Mass Spectrometry at the Cutting Edge

Simultaneous detection of N‐terminal fragment ions in a protein mixture using a ruthenium(II) complex

Distinction of Leu and Ile Using a Ruthenium(II) Complex by MALDI-LIFT-TOF/TOF-MS Analysis

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