非变性质谱和紫外激光解离在蛋白质结构和相互作用分析中的应用OA北大核心CSTPCDMEDLINE

Dynamic changes in the structures and interactions of proteins are closely correlated with their biological functions.However,the precise detection and analysis of these molecules are challenging.Native mass spectrometry(nMS)introduces proteins or protein complexes into the gas phase by electrospray ionization,and then performs MS analysis under near-physiological conditions that preserve the folded state of proteins and their complexes in solution.nMS can provide information on stoichiometry,assembly,and dissociation constants by directly determi-ning the relative molecular masses of protein complexes through high-resolution MS.It can also integrate various MS dissociation technologies,such as collision-induced dissociation(CID),surface-induced dissociation(SID),and ultraviolet photodissociation(UVPD),to analyze the conformational changes,binding interfaces,and active sites of protein complexes,thereby revealing the relationship between their interactions and biological functions.UVPD,especially 193 nm excimer laser UVPD,is a rapidly evolving MS dissociation method that can directly dis-sociate the covalent bonds of protein backbones with a single pulse.It can generate different types of fragment ions,while preserving noncovalent interactions such as hydrogen bonds within these ions,thereby enabling the MS analysis of protein structures with single-amino-acid-site resolution.This review outlines the applications and recent progress of nMS and UVPD in protein dynamic structure and interaction analyses.It covers the nMS techniques used to ana-lyze protein-small-molecule ligand interactions,the structures of membrane proteins and their complexes,and protein-protein interactions.The discussion on UVPD includes the analysis of gas-phase protein structures and interactions,as well as alterations in protein dynamic struc-tures,and interactions resulting from mutations and ligand binding.Finally,this review describes the future development prospects for protein analysis by nMS and new-generation advanced extreme UV light sources with higher brightness and shorter pulses.