NMR and mutational identification of the collagen-binding site of the chaperone Hsp47.
NMR and mutational identification of the collagen-binding site of the chaperone Hsp47.
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DOI：
10.1371/journal.pone.0045930
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Kato K
中科院分区：
文献类型：
--
作者：
Yagi-Utsumi M;Yoshikawa S;Yamaguchi Y;Nishi Y;Kurimoto E;Ishida Y;Homma T;Hoseki J;Nishikawa Y;Koide T;Nagata K;Kato K
热休克蛋白47(Hsp47)作为客户特异性的胶原蛋白伴侣，在胶原蛋白成熟和随后的胚胎发育中起着至关重要的作用。此外，这种蛋白可以成为治疗纤维化的潜在靶点。尽管Hsp47具有重要的生理和病理意义，但目前从结构角度对Hsp47的胶原结合方式知之甚少。在这里，我们描述了一项核磁共振研究，该研究是为了确定Hsp47的胶原结合部位。我们使用了鸡Hsp47，它比人的Hsp47具有更高的溶解度，并针对其色氨酸和组氨酸残基应用了选择性的15N标记方法。根据单个残基的定点突变进行光谱指认。通过检查与三聚胶原肽相互作用时观察到的光谱变化和突变数据，我们成功地将胶原结合位点定位在B/Cβ-Barrel结构域和基于蛇针折叠的3D同源模型中的邻近环。突变分析证实了这一结论。我们的发现为以Hsp47和前胶原相互作用为靶点的化合物的设计提供了分子基础，作为治疗纤维化疾病的药物。
Heat shock protein 47 (Hsp47) acts as a client-specific chaperone for collagen and plays a vital role in collagen maturation and the consequent embryonic development. In addition, this protein can be a potential target for the treatment of fibrosis. Despite its physiological and pathological importance, little is currently known about the collagen-binding mode of Hsp47 from a structural aspect. Here, we describe an NMR study that was conducted to identify the collagen-binding site of Hsp47. We used chicken Hsp47, which has higher solubility than its human counterpart, and applied a selective 15N-labeling method targeting its tryptophan and histidine residues. Spectral assignments were made based on site-directed mutagenesis of the individual residues. By inspecting the spectral changes that were observed upon interaction with a trimeric collagen peptide and the mutational data, we successfully mapped the collagen-binding site in the B/C β-barrel domain and a nearby loop in a 3D-homology model based upon a serpin fold. This conclusion was confirmed by mutational analysis. Our findings provide a molecular basis for the design of compounds that target the interaction between Hsp47 and procollagen as therapeutics for fibrotic diseases.
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影响因子：
--
作者：
Yagi-Utsumi M;Matsuo K;Yanagisawa K;Gekko K;Kato K
通讯作者：
Kato K
影响因子：
15.9
作者：
MASUDA, H;FUKUMOTO, M;NAGATA, K
通讯作者：
NAGATA, K
DOI：
10.1083/jcb.150.6.1499
发表时间：
2000-09-18
期刊：
The Journal of cell biology
影响因子：
--
作者：
Nagai N;Hosokawa M;Itohara S;Adachi E;Matsushita T;Hosokawa N;Nagata K
通讯作者：
Nagata K
影响因子：
2.7
作者：
Koide, T;Nishikawa, Y;Takahara, Y
通讯作者：
Takahara, Y
影响因子：
2.7
作者：
Takeuchi, Koh;Ng, Elise;Wagner, Gerhard
通讯作者：
Wagner, Gerhard