用于细胞和体内成像的结合了对叠氮基-L-苯丙氨酸的位点特异性蛋白质缀合物。
Site-specific protein conjugates incorporating Para-Azido-L-Phenylalanine for cellular and in vivo imaging.
机构信息
Department of Chemistry, Temple University, 1901 N. 13(th) Street, Philadelphia, PA 19122, USA.
Department of Chemistry, Temple University, 1901 N. 13(th) Street, Philadelphia, PA 19122, USA.
出版信息
Methods. 2023 Nov;219:95-101. doi: 10.1016/j.ymeth.2023.10.001. Epub 2023 Oct 5.
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 expressed in 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 for in vitro and in vivo imaging applications based on the representative proteins PTEN and PD-L1. This approach would help elucidate the cellular or in vivo biological activities of the POI.
这项工作的特点是使用琥珀酸抑制介导的非天然氨基酸（UAA）掺入蛋白质，用于各种成像目的。UAA，即对叠氮基-L-苯丙氨酸（pAzF）的定点掺入，提供了一个叠氮基团，可以用于完成应变促进的叠氮-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中或体外表达的蛋白质上进行，从而赋予蛋白质缀合物同质池。提供了一种用于定点蛋白质标记方法的 UAA 掺入的通用程序，允许基于代表性蛋白质 PTEN 和 PD-L1 进行体外和体内成像应用。这种方法将有助于阐明 POI 的细胞或体内生物学活性。
Zotero 插件