A5091300536- Microbial Natural Products and Biosynthesis
- Chemical Thermodynamics and Molecular Structure
- Biochemical effects in animals
- Microbial Metabolic Engineering and Bioproduction
- Advanced Electron Microscopy Techniques and Applications
- Molecular spectroscopy and chirality
- Enzyme Structure and Function
- Electron and X-Ray Spectroscopy Techniques
- Plant biochemistry and biosynthesis
- Genomics and Phylogenetic Studies
- Viral gastroenteritis research and epidemiology
- Animal Virus Infections Studies
- Advanced X-ray Imaging Techniques
- Analytical Chemistry and Chromatography
- HIV Research and Treatment
- CRISPR and Genetic Engineering
- VLSI and Analog Circuit Testing
- Particle Accelerators and Free-Electron Lasers
- Biochemical and Molecular Research
- Chemical Synthesis and Analysis
- Crystal structures of chemical compounds
- Protein Structure and Dynamics
- Crystallization and Solubility Studies
- Enzyme Catalysis and Immobilization
- Plant-Microbe Interactions and Immunity
New England Biolabs (United States)
2023-2024
National Institute of Environmental Health Sciences
2018-2022
National Institutes of Health
2018-2021
University of Michigan
2016-2019
Washtenaw Community College
2018
Michigan United
2016
Abstract Tomographic reconstruction of cryopreserved specimens imaged in an electron microscope followed by extraction and averaging sub-volumes has been successfully used to derive atomic models macromolecules their biological environment. Eliminating biochemical isolation steps required other techniques, this method opens up the cell in-situ structural studies. However, need compensate for errors targeting introduced during mechanical navigation specimen significantly slows down...
We show here that computer game players can build high-quality crystal structures. Introduction of a new feature into the Foldit allows to and real-space refine structures electron density maps. To assess usefulness this feature, we held crystallographic model-building competition between trained crystallographers, undergraduate students, automatic algorithms. After removal disordered residues, team achieved most accurate structure. Analysing target protein competition, YPL067C, uncovered...
Abstract Golden Gate Assembly is a flexible method of DNA assembly and cloning that permits the joining multiple fragments in single reaction through predefined connections. The depends on cutting using Type IIS restriction enzyme, which cuts outside its recognition site therefore can generate overhangs any sequence while separating from generated fragment. By choosing compatible fusion sites, defined order reaction. Conventionally, this has been used to join five eight round, with yield...
Polyketide metabolites produced by modular type I polyketide synthases (PKS) acquire their chemical diversity through the variety of catalytic domains within modules pathway. Methyltransferases are among least characterized common to PKS systems. We determined domain boundaries and activity a C-methyltransferase (C-MT) from curacin A biosynthetic The C-MT catalyzes S-adenosylmethionine-dependent methyl transfer α-position β-ketoacyl substrates linked acyl carrier protein (ACP) or...
Abstract Avian infectious bronchitis is an acute respiratory disease of poultry particular concern for global food security. Investigation Infectious Bronchitis Virus (IBV), the causative agent avian bronchitis, via reverse genetics enables deeper understanding virus biology and a rapid response to emerging variants. Classic methods IBV can be time consuming, rely on recombination introduction mutations, and, depending system, subject genome instability unreliable success rates. In this...
Natural product biosynthetic pathways contain a plethora of enzymatic tools to carry out difficult transformations. Here, we discover an unusual mononuclear iron-dependent methyltransferase that acts in the initiation steps apratoxin A biosynthesis (AprA MT1). Fe3+-replete AprA MT1 catalyzes one or two methyl transfer reactions on substrate malonyl-ACP (acyl carrier protein), whereas Co2+, Fe2+, Mn2+, and Ni2+ support only single transfer. homologues exist within "GNAT" (GCN5-related...
Abstract Cahuitamycins are biofilm inhibitors assembled by a convergent nonribosomal peptide synthetase pathway. Previous genetic analysis indicated that discrete enzyme, CahJ, serves as gatekeeper for cahuitamycin structural diversification. Here, the CahJ protein was probed structurally and functionally to guide formation of new analogues mutasynthetic studies. This enabled in vivo production congener through targeted precursor incorporation.
The unusual feature of a t-butyl group is found in several marine-derived natural products including apratoxin A, Sec61 inhibitor produced by the cyanobacterium Moorea bouillonii PNG 5-198. Here, we determine that A formed as pivaloyl acyl carrier protein (ACP) AprA, polyketide synthase (PKS) loading module biosynthetic pathway. AprA contains an inactive "pseudo" GCN5-related N-acetyltransferase domain (ΨGNAT) flanked two methyltransferase domains (MT1 and MT2) differ distinctly sequence....
Avian infectious bronchitis is an acute respiratory disease of poultry particular concern for global food security. Investigation virus (IBV), the causative agent avian bronchitis, via reverse genetics enables deeper understanding biology and a rapid response to emerging variants. Classic methods IBV can be time consuming, rely on recombination introduction mutations, and, depending system, subject genome instability unreliable success rates. In this study, we have applied data-optimized...
ABSTRACT Tomographic reconstruction of cryopreserved specimens imaged in an electron microscope followed by extraction and averaging sub-volumes has been successfully used to derive atomic models macromolecules their biological environment. Eliminating biochemical isolation steps required other techniques, this method opens up the cell in-situ structural studies. However, need compensate for errors targeting introduced during mechanical navigation specimen significantly slows down...
The cover feature picture shows the substrate binding pocket of promiscuous aryl transferase CahJ in complex with salicyl adenylate. This enzyme activates substrates ATP to form adenylate intermediates that are subsequently transferred an carrier protein (ArCP) as first step cahuitamycin biosynthesis. acts a gatekeeper for structural diversification, and function studies flexibility guided creation new congener. More information can be found communication by D. H. Sherman et al. on page 1595...