Probing the functional hotspots inside protein hydrophobic pockets by in situ photochemical trifluoromethylation and mass spectrometry

Lai，Can1,5; Tang，Zhiyao2; Liu，Zheyi1; Luo，Pan1,3,4; Zhang，Wenxiang1; Zhang，Tingting1,5; Zhang，Wenhao1,5; Dong，Zhe2; Liu，Xinyuan2; Yang，Xueming2,3,4; Wang，Fangjun1,3,5

2024

10.1039/d3sc05106d

Chemical Science   影响因子和分区

2041-6520

2041-6539

Due to the complex high-order structures and interactions of proteins within an aqueous solution, a majority of chemical functionalizations happen on the hydrophilic sites of protein external surfaces which are naturally exposed to the solution. However, the hydrophobic pockets inside proteins are crucial for ligand binding and function as catalytic centers and transporting tunnels. Herein, we describe a reagent pre-organization and in situ photochemical trifluoromethylation strategy to profile the functional sites inside the hydrophobic pockets of native proteins. Unbiased mass spectrometry profiling was applied for the characterization of trifluoromethylated sites with high sensitivity. Native proteins including myoglobin, trypsin, haloalkane dehalogenase, and human serum albumin have been engaged in this mild photochemical process and substantial hydrophobic site-specific and structure-selective trifluoromethylation substitutes are obtained without significant interference to their bioactivity and structures. Sodium triflinate is the only reagent required to functionalize the unprotected proteins with wide pH-range tolerance and high biocompatibility. This “in-pocket” activation model provides a general strategy to modify the potential binding pockets and gain essential structural insights into the functional hotspots inside protein hydrophobic pockets.

SCI ; EI

英语

通讯

2-s2.0-85182940461

Scopus

1.CAS Key Laboratory of Separation Sciences for Analytical Chemistry,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian,116023,China
2.Department of Chemistry,College of Science,Southern University of Science and Technology,Shenzhen,518055,China
3.State Key Laboratory of Molecular Reaction Dynamics,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian,116023,China
4.Institute of Advanced Science Facilities,Shenzhen,518107,China
5.University of Chinese Academy of Sciences,Beijing,100049,China

Lai，Can,Tang，Zhiyao,Liu，Zheyi,et al. Probing the functional hotspots inside protein hydrophobic pockets by in situ photochemical trifluoromethylation and mass spectrometry[J]. Chemical Science,2024.

Lai，Can.,Tang，Zhiyao.,Liu，Zheyi.,Luo，Pan.,Zhang，Wenxiang.,...&Wang，Fangjun.(2024).Probing the functional hotspots inside protein hydrophobic pockets by in situ photochemical trifluoromethylation and mass spectrometry.Chemical Science.

Lai，Can,et al."Probing the functional hotspots inside protein hydrophobic pockets by in situ photochemical trifluoromethylation and mass spectrometry".Chemical Science (2024).