Charting the proteomes of organisms with unsequenced genomes by MALDI-quadrupole time-of-flight mass spectrometry and BLAST homology searching

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Item Type:	Article
Title:	Charting the proteomes of organisms with unsequenced genomes by MALDI-quadrupole time-of-flight mass spectrometry and BLAST homology searching
Creators Name:	Shevchenko, A., Sunyaev, S., Loboda, A., Shevchenko, A., Bork, P., Ens, W. and Standing, K.G.
Abstract:	MALDI-quadrupole time-of-flight mass spectrometry was applied to identify proteins from organisms whose genomes are still unknown. The identification was carried out by successively searching a sequence database-first with a peptide mass fingerprint, then with a packet of noninterpreted MS/MS spectra, and finally with peptide sequences obtained by automated interpretation of the MS/MS spectra. A "MS BLAST" homology searching protocol was developed to overcome specific limitations imposed by mass spectrometric data, such as the limited accuracy of de novo sequence predictions. This approach was tested in a small-scale proteomic project involving the identification of 15 bands of gel-separated proteins from the methylotrophic yeast Pichia pastoris, whose genome has not yet been sequenced and which is only distantly related to other fungi.
Keywords:	Algorithms, Amino Acid Sequence, Cell Line, Factual Databases, Fungal Genome, Kidney, Membrane Proteins, Molecular Sequence Data, Peptide Mapping, Pichia, Proteome, Protein Sequence Analysis, Mass, Matrix-Assisted Laser Desorption-Ionization Spectrometry, Trypsin, Animals, Dogs
Source:	Analytical Chemistry
ISSN:	0003-2700
Publisher:	American Chemical Society
Volume:	73
Number:	9
Page Range:	1917-1926
Date:	1 May 2001
Official Publication:	https://doi.org/10.1021/ac0013709
PubMed:	View item in PubMed

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