Enhanced imaging of protein-specific palmitoylation with HCR-based cis-membrane multi-FRET

作者全名："Fu, Yixin; Qian, Husun; Yang, Yujun; Li, Junjie; Xie, Guoming"

作者地址："[Fu, Yixin; Qian, Husun; Yang, Yujun; Li, Junjie; Xie, Guoming] Chongqing Med Univ, Dept Lab Med, Key Lab Lab Med Diagnost Educ, Minist Educ, Chongqing 400016, Peoples R China; [Fu, Yixin] Zunyi Med Univ, Affiliated Hosp, Dept Blood Transfus, Zunyi 563003, Guizhou, Peoples R China"

通信作者："Xie, GM (通讯作者)，Chongqing Med Univ, Dept Lab Med, Key Lab Lab Med Diagnost Educ, Minist Educ, Chongqing 400016, Peoples R China."

来源：TALANTA

ESI学科分类：CHEMISTRY

WOS号：WOS:001063284300001

JCR分区：Q1

影响因子：6.1

年份：2024

卷号：266

期号：　

开始页：　

结束页：　

文献类型：Article

关键词：PD-L1 palmitoylation; FRET; HCR; Imaging

摘要："Palmitoylation plays an important role in modulating protein trafficking, stability, and activity. The major predicament in protein palmitoylation study is the lack of specific and sensitive tools to visualize protein-specific palmitoylation. Although FRET approach was explored by metabolically labeled palmitic acid and antibody recognized target protein. The trans-membrane strategy suffers from low FRET efficiency due to the donor and acceptor located at different sides of membrane. Herein, we proposed a cis-membrane multi-fluorescence resonance energy transfer (multi-FRET) for amplified visualization of specific palmitoylated proteins through metabolic labeling and targeted recognition. The azido-palmitic acid (azido-PA) was metabolically incorporated into cellular palmitoylated proteins, followed by reacting with dibenzylcylooctyne-modified Cy5 (DBCO-Cy5) through copper-free click chemistry. The protein probe was attached to targeted protein by specific peptide recognition, which initiates a hybridization chain reaction (HCR) amplification process. The cis-membrane labeling method enables effective intramolecular donor-acceptor distance and allow to increase FRET efficiency. Simultaneously, HCR amplification triggered multi-FRET phenomenon with significantly improved FRET efficiency. With the superiority, this strategy has achieved the enhanced FRET imaging of palmitoylated PD-L1 and visualizing the palmitoylation changes of on PD-L1 under drug treatment. Furthermore, the established method successfully amplified visualization of PD-L1 palmitoylation in vivo and mice tumor slice. We envision the approach would provide a useful platform to investigate the effects of palmitoylation on the protein structure and function."

基金机构："National Natural Science Foundation of China [81672112, 81972025]; Project of Zunyi Science and Technology Plan [76]"

基金资助正文："This work was supported by the National Natural Science Foundation of China (No. 81672112, 81972025) and the Project of Zunyi Science and Technology Plan [HZ (2021) No.76]."