Straightforward ladder sequencing of peptides using a Lys-N metalloendopeptidase.
We introduce a method for sequencing peptides by mass spectrometry using a metalloendopeptidase that cleaves proteins at the amino side of lysine (Lys-N). When analyzed by electron transfer dissociation (ETD)-based mass spectrometric sequencing, Lys-N-digested peptides that contain a single lysine residue produce spectra dominated by c-type fragment ions, providing simple ladders for sequence determination. This method should be a valuable strategy for de novo sequencing and the analysis of post-translational modifications.
Taouatas
Nadia
N
Biomolecular Mass Spectrometry and Proteomics Group, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Sorbonnelaan 16, 3584 CA Utrecht, The Netherlands.
Drugan
Madalina M
MM
Heck
Albert J R
AJ
Mohammed
Shabaz
S
D002460:Cell Line; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods; D008666:Metalloendopeptidases / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry*; D020539:Sequence Analysis, Protein / Q000379:methods*
D010455:Peptides; D008666:Metalloendopeptidases; D008239:Lysine