Site-specific protein conjugates incorporating Para-Azido-L-Phenylalanine for cellular and in vivo imaging.
Site-specific protein conjugates incorporating Para-Azido-L-Phenylalanine for cellular and in vivo imaging.
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包含对叠氮基-L-苯丙氨酸的位点特异性蛋白质缀合物，用于细胞和体内成像。
DOI：
10.1016/j.ymeth.2023.10.001
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Wang,RongshengE
中科院分区：
文献类型：
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作者：
Lightle,HaileyE;Kafley,Parmila;Lewis,ToddR;Wang,RongshengE
这项工作的特点是使用琥珀抑制介导的非天然氨基酸（UAA）结合到蛋白质中，用于各种成像目的。对叠氮化物- l -苯丙氨酸（pAzF）的位点特异性结合提供了一个叠氮化物处理，可用于完成菌株促进叠氮化物-炔点击环加成（SPAAC）反应，从而在感兴趣的蛋白质（POI）上引入成像模式，如荧光团或正电子发射断层扫描（PET）示踪剂。这种方法可以直接在哺乳动物细胞系或体外表达的蛋白质上进行，从而获得均匀的蛋白质偶联物库。提供了一种用于UAA掺入的通用程序，用于位点特异性蛋白标记方法，允许基于代表性蛋白PTEN和PD-L1的体外成像应用。这种方法将有助于阐明POI的细胞或体内生物学活性。
This work features the use of amber suppression-mediated unnatural amino acid (UAA) incorporation into proteins for various imaging purposes. The site-specific incorporation of the UAA, p-azido-L-phenylalanine (pAzF), provides an azide handle that can be used to complete the strain promoted azide-alkyne click cycloaddition (SPAAC) reaction to introduce an imaging modality such as a fluorophore or a positron emission tomography (PET) tracer on the protein of interest (POI). Such methodology can be pursued directly in mammalian cell lines or on proteins expressedin vitro, thereby conferring a homogeneous pool of protein conjugates. A general procedure for UAA incorporation to use with a site-specific protein labeling method is provided allowing forin vitroandin vivoimaging applications based on the representative proteins PTEN and PD-L1. This approach would help elucidate the cellular orin vivobiological activities of the POI.