高效液相色谱_荧光检测法测定敬钊毒素_I的磷脂膜结合活性.pdfVIP

2007 11 Vol. 25 N o. 6 Novem ber 2007 Ch inese J ourna l of Chrom atog rap hy 825~ 829 I 1 1, 2 1 曾雄智 , 皮建辉 , 梁宋平 1. , 410081; 2. , 418008 : JZTX , JZTX, JZTX , JZTX 64 47 nm 74 80 nm , JZTX , , JZTX : ; ; ; : O658 : A : 100087 13 2007 0座机电话号码 : Determ ination of JingzhaotoxinIPhospholipidMembrane Binding Activities byH igh Performance Liq id Chromatography with Fl orescence Detection 1 1, 2 1 ZENG X iongzh i , P Jianhu i , L ANG Songp ing 1.T he K ey La bor a tor y of Pr otein Chem i tr y and D evelopm en ta l B io logy ofM in i tr y of Edu ca tion, Hun an N orm a lU n iv er ity, Chan g ha 410081, China ; 2. D epa r tm en t of B ioen gin eer in g, Hua ihua College, Hu a ihua 4180 08, Ch in a Abstract: Jingzhaotoxin JZTX , a 33residue polypep tide w ith three disulfide bonds, w as a novel sp ider neurotox in preferentially inh ib iting cardiac sodium channe l inactivation. ts activities of phospholipid m em braneb inding were studied by a com bination of rev ersedphase high per form an ce liquid chrom atography HPLC and fluorescence spectroscopy. Sm all un ilam e llar v esi cles b inding assays showed that the partition ing of JZTX into lipid b ilayer did not requ ire nega tively charged phospholipids. Further, JZTX also exh ibited a b lue sh ift of 64 nm or 47 nm as well as rededge excitation sh ift of 74 nm or 80 nm in the presence of 75% 1paml itoy l2oleoy l n glycero3phosphoethanolam ine POPE / 25% 1paml itoyl2oleoyln glycero3[ phospho ra c 1g lycerol ] sodium salt POPG or 100% 1paml itoyl2oleoyln glycero3phosphocho line POPC vesicles respectively, suggesting that som e tryptophan residues on the hydrophob ic surface of the tox in were located w ith in a motion restricted m em brane interfacial region. F luo rescence quench ing expermi ents suggested that some tryp tophan residues of JZTX were posi tioned w ith in the m em brane and protected from aqueous quench ing agents. These findings should provide furth