Molecular cloning and expression of Caenorhabditis elegans ERp57-homologue with transglutaminase activity.
Formation of cross-linking between proteins via a gamma-glutamyl-epsilon-lysine residue is an important process in many biological phenomena including apoptosis. Formation of this linkage is catalyzed by the enzyme transglutaminase, which is widely distributed from bacteria to the animal kingdom. The simple multi-cellular organism Caenorhabditis elegans also possesses transglutaminase activity associated with apoptosis [Madi, A. et al. (1998) Eur. J. Biochem. 253, 583-590], but no gene with significant homology to vertebrate or bacterial transglutaminases has been found in the C. elegans genome sequence database. On the other hand, protein disulfide isomerases were recently recognized as a new family of transglutaminases [Chandrashekar, R. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 531-536]. To identify the molecule with transglutaminase activity in C. elegans, we isolated from C. elegans a gene homologous to ERp57, which encodes a protein disulfide isomerase, expressed it in recombinant form, and characterized the transglutaminase and protein disulfide isomerase activities of the resultant protein. The C. elegans ERp57 protein had both enzyme activities, and the transglutaminase activity had similar characteristics to the activity in lysate of the whole worm. These results suggested that the ERp57 homologue was one of the substances with transglutaminase activity in C. elegans.
Natsuka
S
S
Department of Applied Biology, Faculty of Textile Science, Kyoto Institute of Technology, Kyoto 606-8585, Japan. natsuka@ipc.kit.ac.jp
Takubo
R
R
Seki
R
R
Ikura
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D018509:DNA, Helminth / Q000032:analysis; D004926:Escherichia coli / Q000235:genetics; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D007535:Isomerases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D017386:Sequence Homology, Amino Acid; D011503:Transglutaminases / Q000378:metabolism*; D015003:Yeasts
D018509:DNA, Helminth; D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D011503:Transglutaminases; D007535:Isomerases; D019704:Protein Disulfide-Isomerases; C085868:PDIA3 protein, human