De novo peptide sequencing

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Schematic structure of a tandem mass spectrometer.

The de novo -Peptidsequenzierung is a biochemical method for determining the amino acid sequence of proteins or peptides by tandem mass spectrometry (MS / MS). It is a method of protein sequencing .

properties

In comparison to the usual mass spectrometric methods for protein characterization , the de novo peptide sequencing involves a second fragmentation of the peptides. This additional fragmentation can occur in a reflectron e.g. B. can be achieved by a collision-induced (CID) or by a LASER -induced Post-Source Decay (PSD). In the case of multiply charged fragments, further fragmentation can also be achieved by electron capture dissociation or electron impact ionization . In the spectrum, different fragments of a few amino acids are obtained from each fragment of the first round, which by lining up on the basis of the overlaps result in the peptide sequence. This allows the structure of the peptide fragments to be determined in the sense of an N-terminal peptide sequencing. Since this mass spectrometric method does not rely on information about the amino acid sequence, it is called de novo .

Individual evidence

  1. ↑ a b K. F. Medzihradszky: Peptide sequence analysis. In: Methods in enzymology. Volume 402, 2005, ISSN  0076-6879 , pp. 209-244, doi : 10.1016 / S0076-6879 (05) 02007-0 , PMID 16401511 .
  2. ↑ J. Seidler, N. Zinn, ME Boehm, WD Lehmann: De novo sequencing of peptides by MS / MS. In: Proteomics. Volume 10, Number 4, February 2010, ISSN  1615-9861 , pp. 634-649, doi : 10.1002 / pmic.200900459 , PMID 19953542 .
  3. ↑ JP Reilly: Ultraviolet photofragmentation of biomolecular ions. In: Mass spectrometry reviews. Volume 28, number 3, 2009, ISSN  1098-2787 , pp. 425-447, doi : 10.1002 / mas.20214 , PMID 19241462 .
  4. ↑ JW Morgan, JM Hettick, DH Russell: Peptide sequencing by MALDI 193-nm photodissociation TOF MS. In: Methods in enzymology. Volume 402, 2005, ISSN  0076-6879 , pp. 186-209, doi : 10.1016 / S0076-6879 (05) 02006-9 , PMID 16401510 .
  5. ↑ J. Allmer: Algorithms for the de novo sequencing of peptides from tandem mass spectra. In: Expert review of proteomics. Volume 8, number 5, 2011, ISSN  1744-8387 , pp. 645-657, doi : 10.1586 / epr.11.54 , PMID 21999834 .