Autoproteolytic stability of a trypsin from the marine crab Cancer pagurus.
Autoproteolytic stability is a crucial factor for the application of proteases in biotechnology. In contrast to vertebrate enzymes, trypsins from shrimp and crayfish are known to be resistant against autolysis. We show by characterisation of a novel trypsin from the gastric fluid of the marine crab Cancer pagurus that this property might be assigned to the entire class of crustaceans. The isolated and cloned crab trypsin (C.p.TryIII) exhibits all characteristic properties of crustacean trypsins. However, its overall sequence identity to other trypsins of this systematic class is comparatively low. The high resistance against autoproteolysis was determined by mass spectrometry, which revealed a low susceptibility of the N-terminal domain towards autolysis. By homology modelling of the tertiary structure, the elevated stability was attributed to the distinctly different pattern of autolytic cleavage sites, which is conserved in all known crustacean trypsin sequences.
Hehemann
Jan-Hendrik
JH
Institute of Biochemistry and Food Chemistry, University of Hamburg, 20146 Hamburg, Germany.
Redecke
Lars
L
Murugaiyan
Jayaseelan
J
von Bergen
Martin
M
Betzel
Christian
C
Saborowski
Reinhard
R
D000595:Amino Acid Sequence; D000818:Animals; D003386:Brachyura / Q000201:enzymology*; D002118:Calcium / Q000737:chemistry; D004795:Enzyme Stability; D013058:Mass Spectrometry; D008967:Molecular Biology; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000378:metabolism; D011487:Protein Conformation; D014357:Trypsin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D010447:Peptide Hydrolases; D014357:Trypsin; D002118:Calcium