Structure-Based Mutagenesis of Phycobiliprotein smURFP for Optoacoustic Imaging.,ACS Chemical Biology

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Structure-Based Mutagenesis of Phycobiliprotein smURFP for Optoacoustic Imaging.
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2019-08-20 , DOI: 10.1021/acschembio.9b00299
Juan Pablo Fuenzalida Werner ₁ , Kanuj Mishra ₁ , Yuanhui Huang _{1,

2} , Paul Vetschera _{1,

2} , Sarah Glasl ₁ , Andriy Chmyrov _{1,

2,

3} , Klaus Richter ₄ , Vasilis Ntziachristos _{1,

2,

3} , Andre C Stiel ₁

Affiliation

Photo- or optoacoustics (OA) imaging is increasingly being used as a non-invasive imaging method that can simultaneously reveal structure and function in deep tissue. However, the most frequent transgenic OA labels are current fluorescent proteins that are not optimized for OA imaging. Thus, they lack OA signal strength, and their absorption maxima are positioned at short wavelengths, thus giving small penetration depths and strong background signals. Here, we apply insights from our recent determination of the structure of the fluorescent phycobiliprotein smURFP to mutate a range of residues to promote the nonradiative decay pathway that generates the OA signal. We identified hydrophobic and aromatic substitutions within the chromophore-binding pocket that substantially increase the intensity of the OA signal and red-shift the absorption. Our results demonstrate the feasibility of structure-based mutagenesis to repurpose fluorescent probes for OA imaging, and they may provide structure-function insights for de novo engineering of transgenic OA probes.

中文翻译：

藻胆蛋白smURFP的基于结构的诱变，用于光声成像。

光声或光声（OA）成像正越来越多地用作可以同时显示深层组织的结构和功能的非侵入性成像方法。但是，最常见的转基因OA标签是当前的荧光蛋白，尚未针对OA成像进行优化。因此，它们缺乏OA信号强度，其吸收最大值位于短波长处，因此穿透深度较小，背景信号较强。在这里，我们将从对荧光藻胆蛋白smURFP的结构的最新确定中得出的见解应用于突变一系列残基，以促进产生OA信号的非辐射衰变途径。我们在发色团结合袋中发现了疏水和芳香取代基，这些取代基大大增加了OA信号的强度并使吸收红移。

更新日期：2019-08-07

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