ABRF-PRG05: de novo peptide sequence determination.
A common request of proteomics core facilities is protein identification. However, in some instances primary sequence information for the protein in question is not present in public databases. In other cases, the amino acid sequence of a protein may differ in some way from the sequence predicted from the gene sequence in a database as a result of gene mutation, gene splicing, and/or multiple posttranslational modifications. Thus, it may be necessary to determine the sequence of one or more peptides de novo in order to identify and/or adequately characterize the protein of interest. The primary goal of this study was to give participating laboratories an opportunity to evaluate their proficiency in sequencing unknown peptides that are not included in any published database. Samples containing 3-6 pmol each of five synthetic peptides with amino acid sequences that were not present in public databases were sent to 106 laboratories. One nonstandard amino acid was present in one of the peptides. From a comparison of the results obtained by different strategies, participating laboratories will be able to gauge their own capabilities and establish realistic expectations for the approaches that can be used for this determination.
Falick
Arnold M
AM
HHMI Mass Spectrometry Laboratory, University of California, Berkeley, CA, USA.
Kowalak
Jeffrey A
JA
Lane
William S
WS
Phinney
Brett S
BS
Turck
Christoph W
CW
Weintraub
Susan T
ST
West
Karen A
KA
Neubert
Thomas A
TA
D001709:Biotechnology; D002851:Chromatography, High Pressure Liquid; D007753:Laboratories; D013058:Mass Spectrometry / Q000379:methods; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D009842:Oligopeptides; D010455:Peptides