Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids.
Intra- and inter-subunit disulfide bond formation is nonessential in adeno-associated viral capsids.
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DOI：
10.1371/journal.pone.0032163
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Asokan A
中科院分区：
文献类型：
--
作者：
Pulicherla N;Kota P;Dokholyan NV;Asokan A
腺相关病毒（AAV）的衣壳蛋白具有5个保守的半胱氨酸残基。AAV血清型2的结构分析揭示Cys 289和Cys 361在每个单体内彼此相邻定位，而Cys 230和Cys 394位于每个亚基的相对边缘上并且在五聚体界面处并置。Cys 482残基位于三聚体区域内的表面环的基部。虽然合理的分子动力学模拟的基础上，内部或亚基间的二硫化物尚未观察到的结构研究。在目前的研究中，我们产生了一组Cys到Ser突变体，以询问AAV衣壳中二硫键形成的可能性。C289 S、C361 S和C482 S突变体在滴度和转导效率方面与野生型AAV相似。然而，具有C230 S或C394 S突变的AAV衣壳蛋白亚基易于在宿主细胞内发生蛋白酶体降解。蛋白酶体抑制部分阻断了突变体衣壳蛋白的降解，但未能拯救感染性病毒粒子。虽然这些结果表明Cys 230/394对是关键的，但发现C394 V突变体是可行的，但相应的C230 V突变体不是。尽管Cys 230和Cys 394残基对AAV衣壳形成的结构贡献的确切性质仍有待确定，但这些结果支持Cys 289/361或Cys 230/394对内的二硫键形成似乎是不必要的这一观点。这些研究代表了理解驱动AAV衣壳组装的亚基间相互作用的作用的重要一步。
The capsid proteins of adeno-associated viruses (AAV) have five conserved cysteine residues. Structural analysis of AAV serotype 2 reveals that Cys289 and Cys361 are located adjacent to each other within each monomer, while Cys230 and Cys394 are located on opposite edges of each subunit and juxtaposed at the pentamer interface. The Cys482 residue is located at the base of a surface loop within the trimer region. Although plausible based on molecular dynamics simulations, intra- or inter-subunit disulfides have not been observed in structural studies. In the current study, we generated a panel of Cys-to-Ser mutants to interrogate the potential for disulfide bond formation in AAV capsids. The C289S, C361S and C482S mutants were similar to wild type AAV with regard to titer and transduction efficiency. However, AAV capsid protein subunits with C230S or C394S mutations were prone to proteasomal degradation within the host cells. Proteasomal inhibition partially blocked degradation of mutant capsid proteins, but failed to rescue infectious virions. While these results suggest that the Cys230/394 pair is critical, a C394V mutant was found viable, but not the corresponding C230V mutant. Although the exact nature of the structural contribution(s) of Cys230 and Cys394 residues to AAV capsid formation remains to be determined, these results support the notion that disulfide bond formation within the Cys289/361 or Cys230/394 pair appears to be nonessential. These studies represent an important step towards understanding the role of inter-subunit interactions that drive AAV capsid assembly.
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影响因子：
2.9
作者：
Dyson, HJ;Jeng, MF;Holmgren, A
通讯作者：
Holmgren, A
DOI：
10.1016/j.str.2008.03.013
发表时间：
2008-07
期刊：
Structure (London, England : 1993)
影响因子：
--
作者：
Ding F;Tsao D;Nie H;Dokholyan NV
通讯作者：
Dokholyan NV
影响因子：
5.6
作者：
Pornillos O;Ganser-Pornillos BK;Banumathi S;Hua Y;Yeager M
通讯作者：
Yeager M
影响因子：
5.4
作者：
Nam, Hyun-Joo;Lane, Michael Douglas;Agbandje-McKenna, Mavis
通讯作者：
Agbandje-McKenna, Mavis
影响因子：
5.6
作者：
Riolobos, L;Reguera, J;Almendral, JM
通讯作者：
Almendral, JM