A5063926315- RNA and protein synthesis mechanisms
- RNA modifications and cancer
- Advanced biosensing and bioanalysis techniques
- RNA Research and Splicing
- DNA and Nucleic Acid Chemistry
- CRISPR and Genetic Engineering
- Click Chemistry and Applications
- Advanced Fluorescence Microscopy Techniques
- Bacterial Genetics and Biotechnology
- Advanced Biosensing Techniques and Applications
- RNA Interference and Gene Delivery
- Molecular Junctions and Nanostructures
- Lipid Membrane Structure and Behavior
- Genomics and Chromatin Dynamics
- T-cell and B-cell Immunology
- Cellular transport and secretion
- Receptor Mechanisms and Signaling
- Chemical Synthesis and Analysis
- Ubiquitin and proteasome pathways
- Immune Cell Function and Interaction
- Gold and Silver Nanoparticles Synthesis and Applications
- Biosensors and Analytical Detection
- Chromosomal and Genetic Variations
- Monoclonal and Polyclonal Antibodies Research
- interferon and immune responses
Tsinghua University
2017-2025
Center for Life Sciences
2017-2025
State Key Laboratory of Membrane Biology
2024
Beijing Advanced Sciences and Innovation Center
2017-2023
China National Petroleum Corporation (China)
2023
Australian National University
2023
Hangzhou City University
2023
University of Pennsylvania
2010-2018
Peking University
2005-2009
State Key Laboratory for Structural Chemistry of Unstable and Stable Species
2006
In both cancer and infections, diseased cells are presented to human Vγ9Vδ2 T through an 'inside out' signalling process whereby structurally diverse phosphoantigen (pAg) molecules sensed by the intracellular domain of butyrophilin BTN3A11-4. Here we show how-in humans alpaca-multiple pAgs function as 'molecular glues' promote heteromeric association between domains BTN3A1 similar BTN2A1. X-ray crystallography studies visualized that engagement with forms a composite interface for direct...
Abstract The µ-opioid receptor (µOR) is an important target for pain management 1 and molecular understanding of drug action on µOR will facilitate the development better therapeutics. Here we show, using double electron–electron resonance single-molecule fluorescence energy transfer, how ligand-specific conformational changes translate into a broad range intrinsic efficacies at transducer level. We identify several conformations cytoplasmic face that interconvert different timescales,...
Off-target binding and cleavage by Cas9 pose major challenges in its application. How the conformational dynamics of govern nuclease activity under on- off-target conditions remains largely unknown. Here, using intra-molecular single-molecule fluorescence resonance energy transfer measurements, we revealed that apo, sgRNA-bound, dsDNA/sgRNA-bound forms spontaneously transits among three states, mainly reflecting significant mobility catalytic HNH domain. We also uncovered surprising...
Migrasomes are recently discovered organelles, which formed on the ends or branch points of retraction fibers at trailing edge migrating cells. Previously, we showed that recruitment integrins to site migrasome formation is essential for biogenesis. In this study, found prior formation, PIP5K1A, a PI4P kinase converts into PI(4,5)P
Abstract Advances in structural biology have provided important mechanistic insights into signaling by the transmembrane core of G-protein coupled receptors (GPCRs); however, much less is known about intrinsically disordered regions such as carboxyl terminus (CT), which highly flexible and not visible GPCR structures. The β 2 adrenergic receptor’s (β AR) 71 amino acid CT a substrate for kinases binds β-arrestins to regulate signaling. Here we show that AR directly inhibits basal...
Hybridization of nucleic acids with secondary structure is involved in many biological processes and technological applications. To gain more insight into its mechanism, we have investigated the kinetics DNA hybridization/denaturation via fluorescence resonance energy transfer (FRET) on perfectly matched single-base-mismatched strands. hybridization shows non-Arrhenius behavior. At high temperature, apparent activation energies are negative independent structure. In contrast, when...
Significance Elongation factor G (EF-G) uses energy stored in GTP to catalyze movement of transfer RNAs and messenger RNA the ribosome during translocation step prokaryotic protein synthesis. Using single-molecule polarized fluorescence microscopy, three-dimensional rotational motions individual domains EF-G were directly captured, for first time our knowledge, normal translocation. Our observations strongly imply a hybrid model, which initial steps are unlocking driven by force generated...
Abstract New synthetic strategies that exploited the strengths of both chemoselective ligation and recombinant protein expression were developed to prepare K27 di‐ubiquitins (diUb), which enabled mechanistic studies on molecular recognition K27‐linked Ubs by single‐molecule Förster resonance energy transfer (smFRET) X‐ray crystallography. The results revealed free diUb adopted a compact conformation, whereas upon binding UCHL3, was remodeled an open conformation. isopeptide bond remained...
Significance Liquid–liquid phase separation causes formation of membraneless condensates containing concentrated biomolecules, which play essential roles in diverse cellular processes. Currently, micrometer-scale phase-separated are examined routinely to elucidate their functions and mechanisms details. However, limited by current commonly used methods, the transition process from miscible individual molecules is mostly unknown. Herein, we captured nanoscale condensates, whose size, growth...
Maleimide-cysteine chemistry has been a routine practice for the site-specific labeling of fluorophores to proteins since 1950s. This approach, however, cannot bring out best photon budget fluorophores. Here, we systematically measured Cyanine3/5 dye conjugates via maleimide-thiol and amide linkages by counting total emitted photons at single-molecule level. While brightness signal-to-noise ratios do not change significantly, dyes with thioether exhibit more severe photobleaching than...
Hybridization of single-stranded DNA immobilized on the surface gold nanoparticles (GNPs) into double stranded and its subsequent dissociation ssDNA were investigated. Melting curves rates hybridization measured using fluorescence detection based hybridization-induced change. Two distribution functions, namely state rate distribution, proposed in order to take interfacial heterogeneity account quantitatively analyze data. Reaction activation enthalpies entropies GNPs derived compared with...
During protein synthesis, deacylated transfer RNAs leave the ribosome via an exit (E) site after mRNA translocation. How regulates tRNA dissociation and whether functional linkages between aminoacyl (A) E sites modulate dynamics of synthesis have long been debated. Using single molecule fluorescence resonance energy experiments, we find that, during early cycles elongation, tRNAs are often held in until being allosterically released when next binds to A site. This process is regulated by...
Abstract Homologous recombination (HR) is a primary DNA double-strand breaks (DSBs) repair mechanism. The recombinases Rad51 and Dmc1 are highly conserved in the RecA family; mainly responsible for somatic cells during mitosis while only works meiosis germ cells. This spatiotemporal difference probably due to their distinctive mismatch tolerance HR: does not permit HR presence of mismatches, whereas can tolerate certain mismatches. Here, cryo-EM structures Rad51–DNA Dmc1–DNA complexes...
Cas12a is one of the most commonly used Cas proteins for genome editing and gene regulation. The first key step to fulfill its function search target among numerous nonspecific off-target sites. utilizes one-dimensional diffusion along contour dsDNA efficiently target. However, due a lack structural information transient diffusing complex, residues mediating are unknown. Here, combining single-molecule ensemble assays, we found that interactions between at PAM downstream cause asymmetric...
Abstract CasX (also known as Cas12e), a Class 2 CRISPR-Cas system, shows promise in genome editing due to its smaller size compared the widely used Cas9 and Cas12a. Although structures of CasX–sgRNA–DNA ternary complexes have been resolved uncover distinctive NTSB domain, dynamic behaviors are not well characterized. In this study, we employed single-molecule biochemical assays investigate conformational dynamics two homologs, DpbCasX PlmCasX, from DNA binding target cleavage fragment...
Abstract The cellular ESCRT-III (endosomal sorting complex required for transport-III) and Vps4 (vacuolar protein 4) comprise a common machinery that mediates variety of membrane remodelling events. is essential the function by using energy from ATP hydrolysis to disassemble polymer into individual proteins. Here, we report structures ATP-bound E233Q hexamer its with cofactor Vta1 (vps twenty associated 1) at resolutions 3.9 4.2 Å, respectively, determined electron cryo-microscopy. Six...
Significance DNA glycosylase repairs damage to maintain the genome integrity, and thus it is essential for survival of all organisms. However, remains a long-standing puzzle how diffuses along genomic locate sparse aberrant lesion sites efficiently accurately in containing numerous base pairs. Previously, only high-speed–low-accuracy search mode has been characterized experimentally, while low-speed–high-accuracy undetectable. Here, we observed low-speed AlkD translocating, further dissected...
Natural protein‐protein communications, such as those between transcription factors (TFs) and RNA polymerases/ribosomes, underpin cell‐free biosensing systems operating on the transcription/translation (TXTL) paradigm. However, their deployment in field analysis is hampered by delayed response (hour‐level) complex composition of vitro TXTL systems. For this purpose, we present a de novo‐designed ligand‐responsive artificial communication (LIRAC) redefining connection TFs non‐interacting...
Camera-based single-molecule techniques have emerged as crucial tools in revolutionizing the understanding of biochemical and cellular processes due to their ability capture dynamic with high precision, high-throughput capabilities, methodological maturity. However, stringent requirement photon number per frame limited photons emitted by each fluorophore before photobleaching pose a challenge achieving both temporal resolution long observation times. In this work, we introduce MUFFLE,...
SNARE (soluble N-ethylmaleimide sensitive factor attachment protein receptor) proteins are the minimal machinery required for vesicle fusion in eukaryotes. Formation of a highly stable four-helix bundle consisting motif these proteins, drives vesicle/membrane involved several physiological processes such as neurotransmission. Recycling/disassembly membrane is essential and facilitated by an AAA+ ATPase, (NSF) presence adapter protein, α-SNAP. Here we use single-molecule fluorescence...