A5004742269- Enzyme Structure and Function
- Protein Structure and Dynamics
- Protein purification and stability
- Monoclonal and Polyclonal Antibodies Research
- Enzyme Production and Characterization
- Crystallization and Solubility Studies
- Glycosylation and Glycoproteins Research
- Mass Spectrometry Techniques and Applications
- RNA and protein synthesis mechanisms
- Biochemical and Molecular Research
- Yersinia bacterium, plague, ectoparasites research
- Amino Acid Enzymes and Metabolism
- Advanced biosensing and bioanalysis techniques
- Bacterial Genetics and Biotechnology
- Photosynthetic Processes and Mechanisms
- Advanced Proteomics Techniques and Applications
- Plant biochemistry and biosynthesis
- Computational Drug Discovery Methods
- Immunodeficiency and Autoimmune Disorders
- Advanced Biosensing Techniques and Applications
- Cancer, Hypoxia, and Metabolism
- Biochemical and Structural Characterization
- Plant-based Medicinal Research
- Nanofabrication and Lithography Techniques
- Immune Cell Function and Interaction
Institute for Bioscience and Biotechnology Research
2014-2024
National Institute of Standards and Technology
2012-2023
Advanced Bioscience Laboratories (United States)
2014-2023
Research Institute for Bioscience and Biotechnology
2023
National Institute of Standards
2016-2023
Material Measurement Laboratory
2014-2021
University of Maryland, College Park
2016-2020
University of Maryland, Baltimore
2017-2020
MRC Mitochondrial Biology Unit
2012-2020
Medical Research Council
2012-2020
Abstract Complex I is the first and largest enzyme of respiratory chains in bacteria mitochondria. The mechanism which couples spatially separated transfer electrons to proton translocation complex not known. Here we report five crystal structures T. thermophilus with NADH or quinone-like compounds. We also determined cryo-EM major minor native states complex, differing position peripheral arm. Crystal show that binding compounds (but NADH) leads a related global conformational change,...
Naturally occurring metamorphic proteins have the ability to interconvert from one folded state another through either a limited set of mutations or by way change in local environment. Here, we show designed system that it is possible switch reversibly between two most common monomeric folds employing only temperature changes. We demonstrate latent 3α can be unmasked an α/β-plait topology with single V90T amino acid substitution, populating both forms simultaneously. The equilibrium these...
OpenTox provides an interoperable, standards-based Framework for the support of predictive toxicology data management, algorithms, modelling, validation and reporting. It is relevant to satisfying chemical safety assessment requirements REACH legislation as it supports access experimental data, (Quantitative) Structure-Activity Relationship models, toxicological information through integrating platform that adheres regulatory OECD principles. Initial research defined essential components...
Bacteriophages deploy lysins that degrade the bacterial cell wall and facilitate virus egress from host. When applied exogenously, these enzymes destroy susceptible microbes and, accordingly, have potential as therapeutic agents. The most potent lysin identified to date is PlyC, an enzyme assembled two components (PlyCA PlyCB) specific for streptococcal species. Here structure of PlyC holoenzyme reveals a single PlyCA moiety tethered ring-shaped assembly eight PlyCB molecules....
PlyC, a bacteriophage-encoded endolysin, lyses Streptococcus pyogenes (Spy) on contact. Here, we demonstrate that PlyC is potent agent for controlling intracellular Spy often underlies refractory infections. We show the holoenzyme, mediated by its PlyCB subunit, crosses epithelial cell membranes and clears in dose-dependent manner. Quantitative studies using model establish interacts strongly with phosphatidylserine (PS), whereas interaction other lipids weak, suggesting specificity PS as...
Abstract Adoptive cell therapy (ACT) with tumor-specific T cells can mediate cancer regression. The main target of are neoantigens arising from mutations in self-proteins. Although the majority unique to each patient, and therefore not broadly useful for ACT, some shared. We studied oligoclonal T-cell receptors (TCRs) that recognize a shared neoepitope driver mutation p53 oncogene (p53R175H) presented by HLA-A2. Here we report structures wild-type mutant p53–HLA-A2 ligands, as well three...
ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTEnergetics of folding subtilisin BPN'Philip Bryan, Patrick Alexander, Susan Strausberg, Fredrick Schwarz, Wang Lan, Gary Gilliland, and D. Travis GallagherCite this: Biochemistry 1992, 31, 21, 4937–4945Publication Date (Print):June 1, 1992Publication History Published online1 May 2002Published inissue 1 June 1992https://pubs.acs.org/doi/10.1021/bi00136a003https://doi.org/10.1021/bi00136a003research-articleACS PublicationsRequest reuse...
Abstract To better understand how amino acid sequence encodes protein structure, we engineered mutational pathways that connect three common folds (3α, β−grasp, and α/β−plait). The structures of proteins at high sequence-identity intersections in the (nodes) were determined using NMR spectroscopy analyzed for stability function. generate nodes, encoding a smaller fold is embedded structure an ~50% larger new compatible with two sets native interactions designed. This generates pairs 3α or...
When crystallization screening is conducted many outcomes are observed but typically the only trial recorded in literature condition that yielded crystal(s) used for subsequent diffraction studies. The initial hit was optimized and results of all other trials lost. These missing contain information would be useful an improved general understanding crystallization. This paper provides a report data exchange (XDX) workshop organized by several international large-scale laboratories to discuss...
Abstract The resolution of SARS-CoV-2 replication hinges on cell-mediated immunity, wherein CD8 + T cells play a vital role. Nonetheless, the characterization specificity and TCR composition targeting non-spike protein before after infection remains incomplete. Here, we analyzed recognizing six epitopes from nucleocapsid (N) found that slightly increased frequencies N-recognizing but significantly enhanced activation-induced proliferation compared to uninfected donors. N-specific their...
The cyclic AMP receptor protein (CRP, also called catabolite gene activator or CAP) plays a key role in metabolic regulation bacteria and has become widely studied model allosteric transcription factor. On binding its effector cAMP the N-terminal domain, CRP undergoes structural transition to conformation capable of specific DNA C-terminal domain initiation. crystal structures Escherichia coli (EcCRP) cAMP-bound state, both with without DNA, are known, although structure off state...
The crystal structures of two thermally stabilized subtilisin BPN′ variants, S63 and S88, are reported here at 1.8 1.9 Å resolution, respectively. micromolar affinity calcium binding site (site A) has been deleted (Δ75–83) in these enabling the activity thermostability measurements chelating conditions. Each variants includes mutations known previously to increase calcium-independent addition new stabilizing mutations. eight amino acid replacements: D41A, M50F, A73L, Q206W, Y217K, N218S,...
The Biological Macromolecular Crystallization Database (BMCD) has been a publicly available resource since 1988, providing curated archive of information on crystal growth for proteins and other biological macromolecules. BMCD content recently expanded to include 14 372 entries. continues be freely at http://xpdb.nist.gov:8060/BMCD4. In addition, the software adapted support Java-based Lucene query language, enabling detailed searching over specific parameters, explicit search parameter...
Bacillus subtilisin has been a popular model protein for engineering altered substrate specificity. Although some studies have succeeded in increasing the specificity of subtilisin, they also demonstrate that high is difficult to achieve solely by selective binding. In this paper, we analyze structure and transient state kinetic behavior Sbt160, engineered strongly prefer substrates with phenylalanine or tyrosine at P4 position. As previous studies, measure improvements affinity overall...
Abstract We describe the design, kinetic properties, and structures of engineered subtilisin proteases that degrade active form RAS by cleaving a conserved sequence in switch 2. is signaling protein that, when mutated, drives third human cancers. To generate high specificity for target sequence, site was modified to be dependent on cofactor (imidazole or nitrite) protease sub-sites were create linkage between substrate binding. Selective proteolysis arises from 2-step process wherein...
Abstract The enzyme chorismate lyase (CL) catalyzes the removal of pyruvate from to produce 4‐hydroxy benzoate (4HB) for ubiquinone pathway. In Escherichia coli , CL is monomeric, with 164 residues. We have determined structure product complex by crystallographic heavy‐atom methods and report at 1.4‐Å resolution a fully active double Cys‐to‐Ser mutant 2.0‐Å wild‐type. fold involves 6‐stranded antiparallel β‐sheet no spanning helices novel connectivity. bound internally, adjacent sheet, its...