Specific isolation of N‐terminal fragments from proteins and their high‐fidelity de novo sequencing

peer-reviewed · Rapid Communications in Mass Spectrometry · 2007

peer-reviewed · Rapid Communications in Mass Spectrometry · 2007. Minoru Yamaguchi et al. A new method to determine N-terminal amino acid sequences of multiple proteins at low pmol level by a…

Date	2007-10-30
Type	peer-reviewed
Venue	Rapid Communications in Mass Spectrometry
Publisher	Wiley
Contribution	algorithm
DOI	10.1002/rcm.3215
Citations (OpenAlex)	22
Venue 2-year citedness	1.80

Abstract

A new method to determine N-terminal amino acid sequences of multiple proteins at low pmol level by a parallel processing has been developed. The method contains the following five steps: (1) reduction, S-alkylation and guanidination for targeted proteins; (2) coupling with sulfosucccimidyl-2-(biotinamido)ethyl-1,3-dithiopropionate(sulfo-NHS-SS-biotin) to N(alpha)-amino groups of proteins; (3) digestion of the modified proteins by an appropriate protease; (4) specific isolation of N-terminal fragments of proteins by affinity capture using the biotin-avidin system; (5) de novo sequence analysis of peptides by MALDI-TOF-/MALDI-TOF-PSD mass spectrometry with effective utilization of the CAF (chemically assisted fragmentation) method.1 This method is also effective for N-terminal sequencing of each protein in a mixture of several proteins, and for sequencing components of a multiprotein complex. It is expected to become an essential proteomics tool for identifying proteins, especially when used in combination with a C-terminal sequencing method.

Authors

1. Minoru Yamaguchi · Shimadzu (Japan)
2. Takashi Obama
3. Hiroki Kuyama · Osaka University, The University of Osaka
4. Daisuke Nakayama · Shimadzu (Japan)
5. Eiji Ando · Shimadzu (Japan)
6. Taka‐aki Okamura · Osaka University, The University of Osaka
7. Norikazu Ueyama · Osaka University, The University of Osaka
8. Takashi Nakazawa · Nara Women’s University
9. Shigemi Norioka · Osaka University, The University of Osaka
10. Osamu Nishimura · Shimadzu (Japan), The University of Osaka
11. Susumu Tsunasawa · Osaka University, Shimadzu (Japan), The University of Osaka

Methods and tools

• N-terminal fragment isolation de novo sequencing: Isolates protein N-terminal peptides specifically, through reduction, S-alkylation, guanidination and a cleavable biotin coupling, so several proteins can be N-terminally sequenced in parallel at low picomole level.

Cites (2)

• A method for high-sensitivity peptide sequencing using postsource decay matrix-assisted laser desorption ionization mass spectrometry (1999) crossref
• Error-Tolerant Identification of Peptides in Sequence Databases by Peptide Sequence Tags (1994) crossref

Cited by (5)

• Bromine isotopic signature facilitates de novo sequencing of peptides in free‐radical‐initiated peptide sequencing (FRIPS) mass spectrometry (2015) crossref
• C-terminal de novo sequencing of peptides using oxazolone-based derivatization with bromine signature (2011) crossref
• An effective method for de novo peptide sequencing based on phosphorylation strategy and mass spectrometry (2011) crossref
• Selective isolation of N‐terminal peptides from proteins and their de novo sequencing by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry without regard to unblocking or blocking of N‐terminal amino acids (2008) crossref
• A method for N-terminal de novo sequence analysis of proteins by matrix-assisted laser desorption/ionization mass spectrometry (2008) crossref

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