Curcin, a ribosome-inactivating protein with a molecular weight of about 28.2 kD, which stronglyinhibits the protein synthesisin rabbit reticulocytelysate system with anIC50value of about (0.19 ± 0.01) nmol/L, was purified from the seeds of
Jatropha curcas
L. The protein has the activity of rRNA N-glycosidase. Degenerate primers were designed based on the N-terminal partial sequence from purified curcin. The full-length curcin cDNA by RT-PCR and 5'-RACE was cloned. The deduced amino acids sequence indicates that a preprotein with 293 amino acid residues is first translated and then processed to a mature protein with 251 amino acids. The deduced amino acids sequence shares homology of 33% and 57% to those of type I ribosome-inactivating proteins (RIPs) and A chain of type II RIPs, respectively. The sequence encoding mature curcin was integrated into the pQE-30 vector for expression in
Escherichia coil strain M15 (pREP4). The purified recombinant curcin was able to inhibit protein synthesis in rabbit reticulocyte lysate system.
