Boisvert J, Yamamoto T
Can J Microbiol. 1977 Mar;23(3):240-52. doi: 10.1139/m77-036.
Vaccinia virus particles were dissociated into their constituent polypeptides and analysed by sodium dodecyl sulfate (SDS) gel electrophoresis. Thirty-three distinct polypeptide bands were identified and their molecular weights ranged between 11 000 and 150 000 daltons. Specific staining of gels containing polypeptides of dissociated virions revealed the presence of eight glycopeptides. No lipopeptides were detected. Analysis of chemical extracts (urea, guanidine hydrochloride, and alkali treatment) of the virus by SDS gel electrophoresis indicated that a total of 10 to 14 different polypeptides ranging in molecular weights from 11 000 to 70 000 daltons were solubilized. Analysis of detergent extracts and of the remains of extracted viral particles has shown that the detergent Nonidet P-40 (NP-40) solubilized a total of 11 polypeptides of which 6 were glycopeptides. The other detergents sodium deoxycholate (SDC) and cetyl trimethyl ammonium bromide (CTAB) were not as selective. both solubilizing more than 25 of the polypeptides composing the virus. Gel electrophoresis results also indicated that most of the small molecular weight (11 000-70 000 daltons) polypeptides were readily solubilized by NP-40, SDC, and CTAB, while those with molecular weights of 70 000 daltons and higher were not well solubilized. The effects of detergents were also analysed by electron microscopy. Evidence was obtained for subpopulations of viral particles having different susceptibility to detergent extraction.
痘苗病毒颗粒被解离成其组成多肽,并通过十二烷基硫酸钠(SDS)凝胶电泳进行分析。鉴定出33条不同的多肽带,其分子量在11000至150000道尔顿之间。对含有解离病毒粒子多肽的凝胶进行特异性染色,发现存在8种糖肽。未检测到脂肽。通过SDS凝胶电泳对病毒的化学提取物(尿素、盐酸胍和碱处理)进行分析表明,共溶解了10至14种不同的多肽,分子量范围为11000至70000道尔顿。对去污剂提取物和提取后病毒颗粒的残留物进行分析表明,去污剂Nonidet P - 40(NP - 40)共溶解了11种多肽,其中6种是糖肽。其他去污剂脱氧胆酸钠(SDC)和十六烷基三甲基溴化铵(CTAB)的选择性较差,两者都溶解了构成病毒的25种以上的多肽。凝胶电泳结果还表明,大多数小分子量(11000 - 70000道尔顿)的多肽很容易被NP - 40, SDC和CTAB溶解,而分子量在70000道尔顿及以上的多肽则不易溶解。还通过电子显微镜分析了去污剂的作用。获得的证据表明,病毒颗粒亚群对去污剂提取具有不同的敏感性。