[Purification and partial characterization of luffin P1, a peptide with translational inhibitory activity and trypsin inhibitory activity, from seeds of Luffa cylindrica].

Abstract

A peptide, luffin P1, from seeds of Luffa cylindrica, was purified by ammonia sulfate precipitation, CM-52 ion exchange chromatography, Blue-gel affinity chromatography and FPLC Mono S ion exchange chromatography. Its molecular weight was 5226.5 as determined by MALDI-TOF-MS analysis. The sequence of N-terminal 11 amino acids of luffin P1 was identical with the partial N-terminal sequence (from G3 to R13) of 6.5K Arg/Glu rich peptide, which was also isolated from the seeds of Luffa cylindrica. Besides, luffin P1 had a very high homology with a trypsin inhibitor, named C2 peptide, from pumpkin seeds. Interestingly, the purified luffin P1 not only showed a strong inhibitory activity on protein synthesis in rabbit reticulocyte lysate cell-free translation system with IC(50) of 0.6 nmol/L, but also had trypsin inhibitory activity with IC(50) of 22 micromol/L.