Structure of chymotrypsin variant B from Atlantic cod, Gadus morhua.
The amino-acid sequence of chymotrypsin variant B isolated from the pyloric caeca of Atlantic cod has been elucidated. The characterization of the primary structure is based on N-terminal Edman degradation and mass spectrometry of the native protein and enzymatically derived peptides. Chymotrypsin variant B showed 72% sequence identity with the A-variant and 64% and 62%, respectively, with the bovine counterparts A and B, all consisting of 245 amino acids. This new sequence contains a higher proportion of charged residues compared with bovine chymotrypsin but fewer polar hydrogen-bond forming residues which might contribute to its lower thermostability. It also shares the emerging characteristics of other fish serine proteinases which have relatively higher methionine content, including a conserved Met-134 in a loop leading into a domain-connecting strand. The inherent mobility in methionine side-chains may contribute to the maintenance of flexibility at low temperatures. Several amino-acid sequence differences adjacent to the catalytic site are observed in the two cod chymotrypsin variants which also differ in kinetic properties. Unlike the mammalian chymotrypsins, which contain several autolysis sites, cod variant B only contains a single autolysis site. The three-dimensional structures of the A- and B-variants of cod has been modelled on the known crystal structure of bovine alpha-chymotrypsin showing almost superimposable structures.
Leth-Larsen
R
R
Department of Molecular Biology, Odense University, Denmark.
Asgeirsson
B
B
Thórólfsson
M
M
Nørregaard-Madsen
M
M
Højrup
P
P
D000595:Amino Acid Sequence; D000818:Animals; D001267:Atlantic Ocean; D001665:Binding Sites; D002417:Cattle; D002918:Chymotrypsin / Q000737:chemistry*; D005399:Fishes; D007700:Kinetics; D008715:Methionine / Q000032:analysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid
D010446:Peptide Fragments; D008715:Methionine; D002918:Chymotrypsin