Mass spectrometric sequencing of individual peptides from combinatorial libraries via specific generation of chain-terminated sequences.
Combinatorial peptide libraries are a versatile tool for drug discovery. On-bead assays identify reactive peptides by enzyme-catalyzed staining and, usually, sequencing by Edman degradation. Unfortunately, the latter method is expensive and time-consuming and requires free N termini of the peptides. A method of rapid and unambiguous peptide sequencing by utilizing synthesis-implemented generation of termination sequences with subsequent matrix-assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometric analysis is introduced here. The required capped sequences are determined and optimized for a specific peptide library by a computer algorithm implemented in the program Biblio. A total of 99.7% of the sequences of a heptapeptide library sample could be decoded utilizing a single bead for each spectrum. To synthesize these libraries, an optimized capping approach has been introduced.
Hoffmann
Christian
C
Institute of Organic Chemistry, University of Frankfurt, Marie-Curie-Strasse 11, D-60439 Frankfurt am Main, Germany. .
Blechschmidt
Dierk
D
Krüger
Ralf
R
Karas
Michael
M
Griesinger
Christian
C
D000465:Algorithms; D015699:HIV Envelope Protein gp120 / Q000737:chemistry; D006801:Humans; D008832:Microchemistry; D008863:Microspheres; D009842:Oligopeptides / Q000032:analysis* / Q000138:chemical synthesis; D019151:Peptide Library; D020539:Sequence Analysis, Protein / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D015699:HIV Envelope Protein gp120; D009842:Oligopeptides; D019151:Peptide Library