Dissecting the alternatively folded state of the antibody Fab fragment.
Dissecting the alternatively folded state of the antibody Fab fragment.
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剖析抗体 Fab 片段的交替折叠状态。
DOI：
10.1016/j.jmb.2010.04.032
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发表时间：
2010
影响因子：
5.6
通讯作者：
J. Buchner
中科院分区：
文献类型：
--
作者：
Matthias J. Feige;E. R. Simpson;E. Herold;Alexander Bepperling;K. Heger;J. Buchner
完整的抗体和抗原结合片段（Fab）先前已被证明在低pH下形成交替折叠状态（AFS）。这种状态主要由二级结构相互作用组成，三级结构含量减少。AFS可以通过非天然结构的形成，特别是其高稳定性，与熔融球状态区分开来。在本研究中，在已报告诱导AFS的条件下研究了MAK 33（亚型κ/IgG 1的鼠单克隆抗体）Fab片段的分离结构域。令人惊讶的差异，个别领域的能力，形成AFS的观察，尽管在其天然结构的相似性。所有Fab结构域都能够采用AFS，但只有VH（重链可变结构域）可以检测到大量的三级结构，并且需要不同的条件来诱导AFS。VH，在生理条件下最不稳定的结构域，是最稳定的AFS，但所有的结构域显示出显着的稳定性对热和化学展开在其AFS。AFS的形成被发现一般通过未折叠状态进行，对于大多数域具有相似的速率。两者合计，我们的数据揭示了显着差异的生物物理特性的AFS的个别抗体域，反映了高度同源的天然结构域的变化可能。此外，它们允许从抗体Fab片段的AFS中的特异性寡聚体效应中剖析单个结构域贡献。
Intact antibodies and antigen binding fragments (Fab) have been previously shown to form an alternatively folded state (AFS) at low pH. This state consists primarily of secondary structure interactions, with reduced tertiary structure content. The AFS can be distinguished from the molten globule state by the formation of nonnative structure and, in particular, its high stability. In this study, the isolated domains of the MAK33 (murine monoclonal antibody of the subtype κ/IgG1) Fab fragment were investigated under conditions that have been reported to induce the AFS. Surprising differences in the ability of individual domains to form the AFS were observed, despite the similarities in their native structures. All Fab domains were able to adopt the AFS, but only for VH(variable domain of the heavy chain) could a significant amount of tertiary structure be detected and different conditions were needed to induce the AFS. VH, the least stable of the domains under physiological conditions, was the most stable in the AFS, yet all domains showed significant stability against thermal and chemical unfolding in their AFS. Formation of the AFS was found to generally proceed via the unfolded state, with similar rates for most of the domains. Taken together, our data reveal striking differences in the biophysical properties of the AFS of individual antibody domains that reflect the variation possible for domains of highly homologous native structures. Furthermore, they allow individual domain contributions to be dissected from specific oligomer effects in the AFS of the antibody Fab fragment.
影响因子：
16
作者：
Feige, Matthias J.;Groscurth, Sandra;Marcinowski, Moritz;Shimizu, Yuichiro;Kessler, Horst;Hendershot, Linda M.;Buchner, Johannes
通讯作者：
Buchner, Johannes
影响因子：
5.6
作者：
Cardoso, Rosa M. F.;Brunel, Florence M.;Wilson, Ian A.
通讯作者：
Wilson, Ian A.
影响因子：
2.9
作者：
BOLEN, DW;SANTORO, MM
通讯作者：
SANTORO, MM