Tryptic transpeptidation products observed in proteome analysis by liquid chromatography-tandem mass spectrometry.
Commonly, prior to mass spectrometry based analysis of proteins or protein mixtures, the proteins are subjected to specific enzymatic proteolysis. For this purpose trypsin is most frequently used. However, the process of proteolysis is not unflawed. For example, some side activities of trypsin are known and have already been described in the literature (e.g., chymotryptic activity). Here, we describe the occurrence of transpeptidated peptides during standard proteome analysis using two-dimensional polyacrylamide gel electrophoresis followed by mass spectrometric protein identification. Different types of transpeptidated peptides have been detected. The most frequently observed transpeptidation reaction is N-terminal addition of arginine or lysine to peptides. Furthermore, addition of two amino acids to the N-terminus of a peptide has also been detected. Another transpeptidation that we observed, is combination of two peptides, which were originally located in different regions of the analyzed protein. Currently, the full amount of peptides generated by transpeptidation is not clear. However, it should be recognized that protein information is presently lost as these effects are not detectable with available database search software.
Schaefer
Heike
H
Medical Proteom-Center, Ruhr-University Bochum, Bochum, Germany.
Chamrad
Daniel C
DC
Marcus
Katrin
K
Reidegeld
Kai A
KA
Blüggel
Martin
M
Meyer
Helmut E
HE
D000818:Animals; D001120:Arginine / Q000737:chemistry; D002853:Chromatography, Liquid; D045304:Cytochromes c / Q000737:chemistry; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D007782:Lactoglobulins / Q000737:chemistry; D007908:Lens, Crystalline / Q000378:metabolism; D008239:Lysine / Q000737:chemistry; D013058:Mass Spectrometry; D051379:Mice; D008810:Mice, Inbred C57BL; D009211:Myoglobin / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods; D010455:Peptides / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000494:pharmacology*
D007782:Lactoglobulins; D009211:Myoglobin; D010455:Peptides; D011506:Proteins; D020543:Proteome; D045304:Cytochromes c; D001120:Arginine; D014357:Trypsin; D008239:Lysine