Improvements in protein identification by MALDI-TOF-MS peptide mapping.
A new strategy for identifying proteins in sequence data-bases by MALDI-MS peptide mapping is reported. The strategy corrects for systematic deviations of determined peptide molecular masses using information contained in the opened database and thereby renders unnecessary internal spectrum calibration. As a result, data acquisition is simplified and less error prone. Performance of the new strategy is demonstrated by identification of a set of recombinant, human cDNA expression products as well as native proteins isolated from crude mouse brain extracts by 2-D electrophoresis. Using one set of calibration constants for the mass spectrometric analyses, 20 proteins were identified without applying any molecular weight restrictions, which was not possible without data correction. A sequence database search program has been written that performs all necessary calculations automatically, access to which will be provided to the scientific community in the Internet.
Egelhofer
V
V
Max-Planck-Institute for Molecular Genetics, Berlin, Germany.
Büssow
K
K
Luebbert
C
C
Lehrach
H
H
Nordhoff
E
E
D000818:Animals; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D051379:Mice; D010449:Peptide Mapping / Q000295:instrumentation*; D011506:Proteins / Q000032:analysis*; D019837:Ryanodine Receptor Calcium Release Channel / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018076:DNA, Complementary; D011506:Proteins; D019837:Ryanodine Receptor Calcium Release Channel