Homodimerization of the PAS-B domains of hypoxia-inducible factors.
Homodimerization of the PAS-B domains of hypoxia-inducible factors.
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DOI：
10.1021/jp300525n
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发表时间：
2012-06-14
期刊：
影响因子：
--
通讯作者：
Dyson HJ
中科院分区：
文献类型：
--
作者：
Zhu J;Martinez-Yamout M;Cardoso R;Yan J;Love RA;Grodsky N;Brooun A;Dyson HJ
缺氧诱导转录因子（HIF）的Per-Arnt-Sim（PAS）结构域介导在细胞缺氧情况下诱导的HIF-α形式与组成型HIF-β变体之间的异源二聚体形成。以前对HIF-1α PAS结构域的结构表征受到蛋白质稳定性的限制。利用基于已发表的同源蛋白HIF-2α的PAS-B结构域与伴侣HIF-β（也称为ARNT）复合的晶体结构的同源性建模，我们鉴定了具有改善的溶解性、单分散性和稳定性的HIF-1α变体。HIF-1α和HIF-2α的PAS-B结构域的纯化溶液在同二聚体形成的倾向上不同。为了理解这种差异的结构基础，并记录伴随同二聚体形成的结构变化，我们对HIF-1α和HIF-2α的PAS-B结构域以及模拟HIF-2α行为的HIF-1α突变体进行了比较NMR研究。所有这些结构域的NMR光谱非常相似，与它们的氨基酸序列的相似性一致。然而，随着浓度的增加，HIF-1α PAS-B结构域形成二聚体的倾向更大，这使我们能够确定同源二聚体的位点，并指出HIF-1α中可能的序列变化可能会阻碍同源二聚体的形成。
The Per-Arnt-Sim (PAS) domains of hypoxia-inducible transcription factors (HIF) mediate heterodimer formation between the HIF-α forms that are induced in the event of cellular hypoxia and the constitutive HIF-β variants. Previous efforts towards structural characterization of the HIF-1α PAS domains were limited by protein stability. Using homology modeling based on the published crystal structure of the PAS-B domain of the homologous protein HIF-2α in complex with the partner HIF-β (also known as ARNT), we have identified a variant of HIF-1α with improved solubility, monodispersity and stability. Purified solutions of the PAS-B domains of HIF-1α and HIF-2α differ in their propensity for homodimer formation. In an attempt to understand the structural basis for this difference, and to document the structural changes that accompany homodimer formation, we have undertaken a comparative NMR study of the PAS-B domains of HIF-1α and HIF-2α and mutants of HIF-1α that mimic the behavior of HIF-2α. The NMR spectra of all of these domains are very similar, consistent with the similarity of their amino acid sequences. However, the greater propensity of the HIF-1α PAS-B domain to form dimers as the concentration was increased allowed us to determine the site of homodimerization and pointed towards possible sequence changes in HIF-1α that might discourage the formation of homodimers.
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DOI：
10.1073/pnas.88.13.5680
发表时间：
1991-07-01
影响因子：
11.1
作者：
SEMENZA, GL;NEJFELT, MK;ANTONARAKIS, SE
通讯作者：
ANTONARAKIS, SE
影响因子：
4.8
作者：
Hogenesch, JB;Chan, WK;Bradfield, CA
通讯作者：
Bradfield, CA
影响因子：
4
作者：
Semenza, GL
通讯作者：
Semenza, GL
影响因子：
2.7
作者：
DELAGLIO, F;GRZESIEK, S;BAX, A
通讯作者：
BAX, A
影响因子：
5.3
作者：
SEMENZA, GL;WANG, GL
通讯作者：
WANG, GL