Photoinduced reconfiguration to control the protein-binding affinity of azobenzene-cyclized peptides.
Photoinduced reconfiguration to control the protein-binding affinity of azobenzene-cyclized peptides.
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DOI：
10.1039/d0tb01189d
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发表时间：
2020-08-26
期刊：
影响因子：
--
通讯作者：
Menegatti S
中科院分区：
文献类型：
--
作者：
Day K;Schneible JD;Young AT;Pozdin VA;Van Den Driessche G;Gaffney LA;Prodromou R;Freytes DO;Fourches D;Daniele M;Menegatti S
下一代生物识别元件（配体）的影响将取决于在复杂环境中远程控制其对目标生物分子的结合活性的能力。与用于调节结合亲和力的常规机制（pH、离子强度或离液剂）相比，光提供更高的准确性和快速性，并且特别适合于不稳定的靶标。在这项研究中，我们展示了一种通用的方法来开发偶氮苯环化的肽配体与目标蛋白的光控制的亲和力。光引发偶氮苯的顺式/反式异构化，这导致环肽从非结合构型到结合构型的主要结构重排。实现这一目标的关键是在低光剂量下实现有效光异构化的能力以及顺式和反式异构体的时间稳定性。我们通过设计靶向血管细胞粘附标志物1（VCAM 1）（一种与干细胞功能有关的细胞标志物）的光开关肽来证明我们的方法。从已知的VCAM 1结合线性肽开始，通过将计算机模拟设计与经由光谱学和表面等离子体共振的实验表征相结合来鉴定具有选择性光控结合的偶氮苯环化变体的集合。变体cycloAZOB[G-VHAKQHRN-K]的特征是快速、光控结合VCAM 1（KD，反式/KD，顺式~ 130）。利用生物素-cycloAZOB [G-VHAKQHRN-K]标记脑微血管内皮细胞（BMEC），显示与顺式构型的抗VCAM 1抗体共定位，反式构型的结合可忽略不计。
The impact of next-generation biorecognition elements (ligands) will be determined by the ability to remotely control their binding activity for a target biomolecule in complex environments. Compared to conventional mechanisms for regulating binding affinity (pH, ionic strength, or chaotropic agents), light provides higher accuracy and rapidity, and is particularly suited for labile targets. In this study, we demonstrate a general method to develop azobenzene-cyclized peptide ligands with light-controlled affinity for target proteins. Light triggers a cis/trans isomerization of the azobenzene, which results in a major structural rearrangement of the cyclic peptide from a non-binding to a binding configuration. Critical to this goal are the ability to achieve efficient photo-isomerization under low light dosage and the temporal stability of both cis and trans isomers. We demonstrated our method by designing photo-switchable peptides targeting vascular cell adhesion marker 1 (VCAM1), a cell marker implicated in stem cell function. Starting from a known VCAM1-binding linear peptide, an ensemble of azobenzene-cyclized variants with selective light-controlled binding were identified by combining in silico design with experimental characterization via spectroscopy and surface plasmon resonance. Variant cycloAZOB[G-VHAKQHRN-K] featured rapid, light-controlled binding of VCAM1 (KD,trans/KD,cis ~ 130). Biotin-cycloAZOB[G-VHAKQHRN-K] was utilized to label brain microvascular endothelial cells (BMECs), showing co-localization with anti-VCAM1 antibodies in cis configuration and negligible binding in trans configuration.
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影响因子：
14
作者：
Aubin H;Nichol JW;Hutson CB;Bae H;Sieminski AL;Cropek DM;Akhyari P;Khademhosseini A
通讯作者：
Khademhosseini A
影响因子：
15
作者：
Bellotto, Silvia;Chen, Shiyu;Heinis, Christian
通讯作者：
Heinis, Christian
影响因子：
6.3
作者：
BERENDSEN, HJC;VANDERSPOEL, D;VANDRUNEN, R
通讯作者：
VANDRUNEN, R
DOI：
10.1165/ajrcmb.18.5.3052
发表时间：
1998-05-01
影响因子：
6.4
作者：
Blease, K;Seybold, J;Burke-Gaffney, A
通讯作者：
Burke-Gaffney, A
影响因子：
3.6
作者：
Bryant, SJ;Nuttelman, CR;Anseth, KS
通讯作者：
Anseth, KS