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  Protein Identification by MALDI-TOF-MS Peptide Mapping: A New Strategy

Egelhofer, V., Gobom, J., Seitz, H., Giavalisco, P., Lehrach, H., & Nordhoff, E. (2002). Protein Identification by MALDI-TOF-MS Peptide Mapping: A New Strategy. Analytical Chemistry, 74(8), 1760-1771.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-8C32-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-8C33-8
Genre: Journal Article

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 Creators:
Egelhofer, Volker1, Author
Gobom, Johan2, Author              
Seitz, Harald2, Author              
Giavalisco, Patrick1, Author
Lehrach, Hans2, Author              
Nordhoff, Eckhard2, Author              
Affiliations:
1Max Planck Society, ou_persistent13              
2Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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 Abstract: A new strategy for identifying proteins by MALDI-TOF-MS peptide mapping is reported. In contrast to current approaches, the strategy does not rely on a good relative or absolute mass accuracy as the criterion that discriminates false positive results. The protein sequence database is first searched for all proteins that match a minimum five of the submitted masses within the maximum expected relative errors when the default or externally determined calibration constants are used, for instance, ±500 ppm. Typically, this search retrieves many thousand candidate sequences. Assuming initially that each of these is the correct protein, the relative errors of the matching peptide masses are calculated for each candidate sequence. Linear regression analysis is then performed of the calculated relative errors as a function of m/z for each candidate sequence, and the standard deviation to the regression is used to distinguish the correct sequence among the candidates. We show that this parameter is independent of whether the mass spectrometric data were internally or externally calibrated. The result is a search engine that renders internal spectrum calibration unnecessary and adapts to the quality of the raw data without user interference. This is made possible by a dynamic scoring algorithm, which takes into account the number of matching peptide masses, the percentage of the protein's sequence covered by these peptides and, as new parameter, the determined standard deviation. The lower the standard deviation, the less cleavage peptides are required for identification and vice versa. Performance of the new strategy is demonstrated and discussed. All necessary computing has been implemented in a computer program, free access to which is provided in the Internet.

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Language(s): eng - English
 Dates: 2002-03-16
 Publication Status: Published in print
 Pages: -
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 Rev. Method: -
 Identifiers: eDoc: 24742
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Title: Analytical Chemistry
Source Genre: Journal
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Pages: - Volume / Issue: 74 (8) Sequence Number: - Start / End Page: 1760 - 1771 Identifier: -