Semenova N P, Prokudina E P, Chumakov V M, Zuev E A, Zolotareva I L
Mol Gen Mikrobiol Virusol. 1995 Jul-Sep(3):43-8.
Two high-molecular-weight types of in vivo formed nucleoproteins (NP) capable of reacting with anti-NP monoclonal antibodies were detected in continuous MDCK cells and primary CEFs, along with the monomeric virus NP. High-molecular NP were stable at 37 degrees C, despite the presence of dodecyl sulfate and B-mercaptoethanol, but were converted to monomeric NP at 37 degrees C in the presence of 6 M urea, 1 M NaCl, and at acid pH. Hence, high-molecular-weight NP may be the polymeric forms of NP, which are probably stabilized by noncovalent bonds. The kinetics of NP-polymers formation suggests that NP-monomers are their precursors and NP-polymers are formed rapidly after monomeric NP molecules synthesis. Both forms of NP-polymers are detected in the sedimented and soluble fractions of not only cell lysate, but in the extracellular maintenance medium as well. NP-polymers are resistant to RNAse and are more resistant to protease than NP-monomers.
在连续传代的MDCK细胞和原代鸡胚成纤维细胞(CEFs)中,除了单体病毒核蛋白(NP)外,还检测到两种能够与抗NP单克隆抗体发生反应的体内形成的高分子量核蛋白(NP)。尽管存在十二烷基硫酸钠和β-巯基乙醇,但高分子量NP在37℃下是稳定的,但在6M尿素、1M氯化钠存在以及酸性pH条件下,37℃时会转化为单体NP。因此,高分子量NP可能是NP的聚合形式,它们可能通过非共价键稳定。NP聚合物形成的动力学表明,NP单体是它们的前体,并且在单体NP分子合成后迅速形成NP聚合物。两种形式的NP聚合物不仅在细胞裂解物的沉淀和可溶部分中被检测到,在细胞外维持培养基中也能检测到。NP聚合物对RNA酶有抗性,并且比NP单体对蛋白酶更具抗性。