A napin-like polypeptide with translation-inhibitory, trypsin-inhibitory, antiproliferative and antibacterial activities from kale seeds.
A heterodimeric napin-like polypeptide with translation-inhibiting and antibacterial activities has been isolated from kale seeds. The purification procedure entailed ion-exchange chromatography on dielthylaminoethyl (DEAE)-cellulose, affinity chromatography on Affi-gel blue gel, ion-exchange chromatography by fast protein liquid chromatography (FPLC) on Mono S, and gel filtration by FPLC on Superdex 75. The napin-like polypeptide was unadsorbed on DEAE-cellulose but adsorbed on Affi-gel blue gel and Mono S. Its 7-kDa large subunit differs in N-terminal amino acid sequence from the 4-kDa small subunit. The polypeptide inhibited translation in the rabbit reticulocyte lysate system with an IC50 of 37.5 nM. This activity was preserved between pH 5 and pH 11, and between 10 and 40 degrees C. It fell to a low level at pH 3 and pH 13 and at 70 degrees C. Antibacterial activity against Bacillus, Megabacterium, and Pseudomonas species and antiproliferative activity against leukemia L1210 cells were observed. However, the polypeptide did not exert antifungal, ribonuclease, or protease activity.
Department of Biochemistry, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
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D055516:2S Albumins, Plant; D000595:Amino Acid Sequence; D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002474:Cell-Free System; D002850:Chromatography, Gel; D004305:Dose-Response Relationship, Drug; D018922:HL-60 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D020128:Inhibitory Concentration 50; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D010940:Plant Proteins / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D017434:Protein Structure, Tertiary; D012260:Ribonucleases / Q000378:metabolism; D013696:Temperature; D014357:Trypsin / Q000737:chemistry* / Q000494:pharmacology
D055516:2S Albumins, Plant; D000890:Anti-Infective Agents; D010455:Peptides; D010940:Plant Proteins; C050363:napin protein, Brassica napus; D012260:Ribonucleases; D014357:Trypsin