A napin-like polypeptide from dwarf Chinese white cabbage seeds with translation-inhibitory, trypsin-inhibitory, and antibacterial activities.
Napins are 1:1 disulfide-linked complexes of a smaller (ca. 4kDa) subunit and a larger (ca. 10kDa) subunit. The intent of the present study was to ascertain the production of napin by the seeds of a Brassica species that has not been examined previously, and also to explore new biological activities of the napin. A heterodimeric 11-kDa napin-like polypeptide has been isolated from Chinese white cabbage (Brassica chinensis cv dwarf) seeds with a protocol comprising ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel, fast protein liquid chromatography (FPLC)-ion exchange chromatography on Mono S and FPLC-gel filtration on Superdex 75. The N-terminal sequence of the 7-kDa subunit manifests striking similarity to napin large chain, albumin and trypsin inhibitor. The N-terminal sequence of the 4-kDa subunit is homologous to napin large chain and an antimicrobial peptide. The napin-like polypeptide inhibited translation in the rabbit reticulocyte system with an IC50 of 18.5nM. This translation-inhibitory activity was stable between pH 4 and 11, and between 10 and 40 degrees C. The polypeptide inhibited trypsin with a higher potency ( IC50 = 8.5 microM) than it inhibited chymotrypsin (IC50 = 220 microM), but was devoid of ribonuclease and antifungal activities. It manifested antibacterial activity against Pseudomonas aeruginosia, Bacillus subtilis, Bacillus cereus, and Bacillus megaterium. The results revealed that the napin-like polypeptide from Chinese white cabbage seeds exhibited some potentially exploitable activities.
Ngai
P H K
PH
Department of Biochemistry, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China.
Ng
T B
TB
D055516:2S Albumins, Plant; D000595:Amino Acid Sequence; D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry*; D001937:Brassica / Q000737:chemistry*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000737:chemistry*; D014176:Protein Biosynthesis; D011817:Rabbits; D012156:Reticulocytes; D012639:Seeds / Q000737:chemistry*; D014357:Trypsin / Q000378:metabolism; D014361:Trypsin Inhibitors / Q000737:chemistry*
D055516:2S Albumins, Plant; D000900:Anti-Bacterial Agents; D010940:Plant Proteins; D014361:Trypsin Inhibitors; C050363:napin protein, Brassica napus; D014357:Trypsin