MAS NMR on a Red/Far-Red Photochromic Cyanobacteriochrome All2699 from Nostoc

MAS NMR on a Red/Far-Red Photochromic Cyanobacteriochrome All2699 from Nostoc
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来自发菜的红色/远红光致变色蓝藻色素 All2699 的 MAS NMR

DOI:
10.3390/ijms20153656
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发表时间:
2019-07
影响因子:
5.6
通讯作者:
Song Chen
Song Chen
中科院分区:
生物学2区
文献类型:
--
作者:
Xu Qian-Zhao;Bielytskyi Pavlo;Otis James;Lang Christina;Hughes Jon;Zhao Kai-Hong;Losi Aba;Gaertner Wolfgang;Song Chen

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与典型的光敏色素不同,蓝细菌色素(CBCR)的GAF结构域可以自主结合胆色素,并且足以进行功能性光循环。尽管CBCR的光谱多样性令人惊讶,但来自蓝细菌Nostoc 7120的三GAF结构域光感受器all 2699的GAF 1结构域是已知的唯一CBCR-GAF,其从红色吸收(Pr)暗态转化为远红色吸收(Pfr)光产物,类似于更保守的光敏色素。在这里,我们报告了一个固态NMR光谱研究的所有2699 g1在其Pr状态。结论性的NMR证据揭示了在四面体C31原子处的特定立体化学异质性,而晶体结构仅显示在该手性中心处的R-立体化学。对包含所有2699的GAF 1和GAF 2结构域的构建体进行额外的NMR实验,显示GAF 1 -2构建体中的发色团-蛋白质相互作用的更高精度。一个3D Pr结构模型的all 2699 g1 -2构建预测的舌状区域从GAF 2域(类似于典型的光敏色素)的方向上的发色团,屏蔽它从溶剂中延伸。此外,这种稳定化元件仅允许发色团-蛋白质键的R-立体化学。在发夹样尖端的三个保守基序上进行的定点突变证实了舌区与GAF 1结合的发色团的相互作用。
Unlike canonical phytochromes, the GAF domain of cyanobacteriochromes (CBCRs) can bind bilins autonomously and is sufficient for functional photocycles. Despite the astonishing spectral diversity of CBCRs, the GAF1 domain of the three-GAF-domain photoreceptor all2699 from the cyanobacterium Nostoc 7120 is the only CBCR-GAF known that converts from a red-absorbing (Pr) dark state to a far-red-absorbing (Pfr) photoproduct, analogous to the more conservative phytochromes. Here we report a solid-state NMR spectroscopic study of all2699g1 in its Pr state. Conclusive NMR evidence unveils a particular stereochemical heterogeneity at the tetrahedral C31 atom, whereas the crystal structure shows exclusively the R-stereochemistry at this chiral center. Additional NMR experiments were performed on a construct comprising the GAF1 and GAF2 domains of all2699, showing a greater precision in the chromophore–protein interactions in the GAF1-2 construct. A 3D Pr structural model of the all2699g1-2 construct predicts a tongue-like region extending from the GAF2 domain (akin to canonical phytochromes) in the direction of the chromophore, shielding it from the solvent. In addition, this stabilizing element allows exclusively the R-stereochemistry for the chromophore-protein linkage. Site-directed mutagenesis performed on three conserved motifs in the hairpin-like tip confirms the interaction of the tongue region with the GAF1-bound chromophore.
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