PrP106—126多肽抑制14—3—3β蛋白二聚体形成的研究  

Study on PrP106-126 pepfide disturbed dimeration of 14-3-3β

作　　者：宋芹芹[1] 孙鹏[2] 宋娟[1] 盛琳君[1] 张瑾[1] 韩俊[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所传染病预防控制所国家重点实验室,北京102206 [2]内蒙古医学院

出　　处：《中华实验和临床病毒学杂志》2013年第2期109-111,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：传染病预防控制国家重点实验室重点项目

摘　　要：目的探讨PrP蛋白和PrP106-126多肽对14—3．3蛋白二聚体形成的影响方法将重组表达纯化的0．25μg、0．5μg和1μgPrP蛋白、人工合成的PrP06—126肽分别与重组的14—3—3β共孵育，分别通过非变性聚丙烯酰胺凝胶的WesternB1ott检测PrP蛋白和PrP106-126肽对14．3．3B二聚体和多聚体的形成影响。将不同浓度的PrP与250μmo1／L的PrP106．126肽和14-3-3蛋白共孵育，检测PrP蛋白对PrP106—126肽对14-3-3β二聚体和多聚体的形成影响。随后，将200g,mo1／L、100μmo1／L、50μmo1／L的PrP106—126多肽分别作用于HeLa细胞8h后，检测细胞内的14-3-3二聚体形成。结果PrP蛋白与14-3—3蛋白共孵育后，14—3—3二聚体和多聚体的信号随着PrP浓度的增加明显增强；PrP106—126多肽共孵育时，14-3—3二聚体和多聚体的信号随着PrP浓度的增加而减弱；不同浓度的PrP蛋白可以拮抗PrP106—126多肽对14-3-3B二聚体的形成。而且PrP106．126多肽可以干扰HeLa细胞中的二聚体形成。结论PrP蛋白促进14—3—3二聚体的形成，而PrP106-126多肽干扰二聚体的形成，且PrP蛋白具有拮抗PrP106-126肽干扰14-3-3二聚体形成的作用。Objective To investigate both PrP and PrP106-126 peptide effect on 14-3-3β dimeration. Methods 14-3-313 were incubated with different does recombinant PrP or PrPI06-126 peptide, both 14-3-313 dimer and polymer were separated 15% non-denaturing polyacrylamide gel electrophoresis （PAGE） and the 14-3-3 dimers were evaluated using 14-3-3β-specific Western blotting. And then,14-3-3β dimeration buffer were incubated with different does recombinant PrP and 250 μmol/L PrP106-126 peptide, 14-3-3β dimer and polymer were detected by above methods. Cellular 14-3-3 dimer were also detected after PrPI06-126 peptide were added to HeLa cell for g hours. Results Recombinant full-length PrP facilitated the dimerization of 14-3-3β and PrP106-126 disturbed 14-3-3β dimeration as both have dose dependence effect. PrP antagonized PrP106-126-induced 14-3-3β dimer with PrP protein increase in vitro. Cellular 14-3-3 dimerization also decreased after treatment of peptide PrP106-126 on HeLa cells for 8 hours. Conculsion Dimerization process of 14-3-3β was promoted by full-length PrP （PrP23-231 ） but inhibited by peptide PrP106-126 in vitro. PrP agonized PrP106-126-induced inhibition of 14-3-3 dimeration. PrP106-126 inhibited cellular 14-3-3 dimerization.

关 键 词：蛋白质工程 肽类 二聚作用 

分 类 号：R[医药卫生]