A5088153235- Advanced Electron Microscopy Techniques and Applications
- Plant nutrient uptake and metabolism
- Bacterial Genetics and Biotechnology
- Mitochondrial Function and Pathology
- RNA Research and Splicing
- Genetic Neurodegenerative Diseases
- Pancreatic function and diabetes
- Protein Structure and Dynamics
- Advanced Fluorescence Microscopy Techniques
- Amino Acid Enzymes and Metabolism
- Heat shock proteins research
- Advanced X-ray Imaging Techniques
- Electron and X-Ray Spectroscopy Techniques
- Enzyme Structure and Function
- RNA and protein synthesis mechanisms
- DNA Repair Mechanisms
- Machine Learning in Bioinformatics
- Genetics and Neurodevelopmental Disorders
- Genomics and Phylogenetic Studies
- Biochemical Analysis and Sensing Techniques
- Cellular transport and secretion
- Cancer Cells and Metastasis
- Connexins and lens biology
- Lipid Membrane Structure and Behavior
- Cell Adhesion Molecules Research
SLAC National Accelerator Laboratory
2020-2024
Stanford Synchrotron Radiation Lightsource
2020-2024
Stanford University
2020-2023
Baylor College of Medicine
2016-2020
Clark University
2020
Superresolution fluorescence microscopy and cryogenic electron tomography (CET) are powerful imaging methods for exploring the subcellular organization of biomolecules. based on covalent labeling highlights specific proteins has sufficient sensitivity to observe single fluorescent molecules, but reconstructions lack detailed cellular context. CET molecular-scale resolution lacks nonperturbative intracellular techniques. Here, we describe an scheme that correlates single-molecule...
Abstract Huntington’s disease (HD) is caused by an expanded CAG repeat in the huntingtin gene, yielding a Huntingtin protein with polyglutamine tract. While experiments patient-derived induced pluripotent stem cells (iPSCs) can help understand disease, defining pathological biomarkers remains challenging. Here, we used cryogenic electron tomography to visualize neurites HD patient iPSC-derived neurons varying repeats, and primary cortical from BACHD, deltaN17-BACHD, wild-type mice. In...
While many 3D structures of cation-coupled transporters have been determined, the mechanistic details governing obligatory coupling and functional regulations still remain elusive. The bacterial melibiose transporter (MelB) is a prototype major facilitator superfamily transporters. With conformation-selective nanobody, we determined low-sugar affinity inward-facing Na + -bound cryoEM structure. available outward-facing sugar-bound showed that N- C-terminal residues inner barrier contribute...
Abstract Although acknowledged to be variable and subjective, manual annotation of cryo-electron tomography data is commonly used answer structural questions create a “ground truth” for evaluation automated segmentation algorithms. Validation such lacking, but critical understanding the reproducibility annotations. Here, we voxel-based similarity scores variety specimens, ranging in complexity segmented by several annotators, quantify variation among their In addition, have identified...
While many 3D structures of cation-coupled transporters have been determined, the mechanistic details governing obligatory coupling and functional regulations still remain elusive. The bacterial melibiose transporter (MelB) is a prototype major facilitator superfamily transporters. With conformation-selective nanobody, we determined low-sugar affinity inward-facing Na+-bound cryoEM structure. available outward-facing sugar-bound showed that N- C-terminal residues inner barrier contribute to...
Abstract Migration and invasion are key properties of metastatic cancer cells. These can be acquired through intrinsic reprogramming processes such as epithelial-mesenchymal transition. In this study, we discovered an alternative “migration-by-tethering” mechanism which cells gain the momentum to migrate by adhering mesenchymal stem or osteoblasts. This tethering is mediated both heterotypic adherens junctions gap junctions, leads a unique cellular protrusion supported cofilin-coated actin...
SUMMARY Three-dimensional (3D) visualization of vitrified cells can uncover structures subcellular complexes without chemical fixation or staining. Here, we present a pipeline integrating three imaging modalities to visualize the same specimen at cryogenic temperature different scales: cryo-fluorescence confocal microscopy, volume cryo-focused ion beam scanning electron and transmission cryo-electron tomography. Our proof-of-concept benchmark revealed 3D distribution organelles in whole...
While many 3D structures of cation-coupled transporters have been determined, the mechanistic details governing obligatory coupling and functional regulations still remain elusive. The bacterial melibiose transporter (MelB) is a prototype Na + -coupled major facilitator superfamily transporters. With conformational nanobody (Nb), we determined low-sugar affinity inward-facing -bound cryoEM structure. Collectively with available outward-facing sugar-bound structures, both outer inner barriers...
Abstract While many 3D structures of cation-coupled transporters have been determined, the mechanistic details governing obligatory coupling and functional regulations still remain elusive. The bacterial melibiose transporter (MelB) is a prototype Na + -coupled major facilitator superfamily transporters. With conformational nanobody (Nb), we determined low-sugar affinity inward-facing -bound cryoEM structure. Collectively with available outward-facing sugar-bound structures, both outer inner...
Electron crystallography has recently gained attentions in multiple fields of research, as it been demonstrated to determine atomic structures for inorganic, organic, and macromolecular materials from nano-sized crystals that were not amenable conventional X-ray crystallography. Here, we demonstrate continuous-rotation microcrystal electron diffraction (microED) a 200 kV transmission microscope using DE-64 camera-a low-noise direct detector can accommodate linear response up ∼1200 electrons...
Abstract Huntington’s Disease (HD) is caused by an expanded CAG repeat in the huntingtin gene, yielding a Huntingtin protein with polyglutamine tract. Patient-derived induced pluripotent stem cells (iPSCs) can help understand disease; however, defining pathological biomarkers challenging. Here, we used cryogenic electron tomography to visualize neurites HD patient iPSC-derived neurons varying repeats, and primary cortical from BACHD, deltaN17-BACHD, wild-type mice. In models, discovered...
While many 3D structures of cation-coupled transporters have been determined, the mechanistic details governing obligatory coupling and functional regulations still remain elusive. The bacterial melibiose transporter (MelB) is a prototype Na + -coupled major facilitator superfamily transporters. With conformational nanobody (Nb), we determined an inward-facing -bound, low-sugar affinity cryoEM structure. It first structure (MFS) with experimentally cation binding, also mimicking...
Abstract Huntington’s Disease (HD) is caused by an expanded CAG repeat in the huntingtin gene, yielding a Huntingtin protein with polyglutamine tract. Patient-derived induced pluripotent stem cells (iPSCs) can help understand disease; however, defining pathological biomarkers challenging. Here, we used cryogenic electron tomography to visualize neurites HD patient iPSC-derived neurons varying repeats, and primary cortical from BACHD, deltaN17-BACHD, wild-type mice. In models, discovered...
Journal Article Cryo-EM Information Management System and Sample Evaluation at Stanford-SLAC Center Get access Htet Khant, Khant Division of CryoEM Bioimaging, Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Menlo Park, CA, USA Search for other works by this author on: Oxford Academic Google Scholar Yee T Li, Li Scientific Computing Services, Murali Shankar, Shankar Corey Hecksel, Hecksel Patrick Mitchell, Mitchell Yan Liu, Liu Lydia-Marie Joubert, Joubert...