A5026956570- RNA and protein synthesis mechanisms
- Metal-Catalyzed Oxygenation Mechanisms
- Ubiquitin and proteasome pathways
- 14-3-3 protein interactions
- RNA modifications and cancer
- Bacterial Genetics and Biotechnology
- Microbial metabolism and enzyme function
- Peroxisome Proliferator-Activated Receptors
- Monoclonal and Polyclonal Antibodies Research
- Click Chemistry and Applications
- Metal complexes synthesis and properties
- Bacterial biofilms and quorum sensing
- Genomics and Phylogenetic Studies
- Chemical Synthesis and Analysis
- Glycosylation and Glycoproteins Research
- DNA and Nucleic Acid Chemistry
- Oral microbiology and periodontitis research
- Microbial Natural Products and Biosynthesis
- Vibrio bacteria research studies
- Advanced biosensing and bioanalysis techniques
- Antimicrobial Peptides and Activities
- Microbial bioremediation and biosurfactants
- Machine Learning in Bioinformatics
- Legionella and Acanthamoeba research
- Porphyrin Metabolism and Disorders
Oregon State University
2012-2024
Corvallis Environmental Center
2021
Cornell University
2013-2019
New York State College of Veterinary Medicine
2014-2016
Institute of Molecular Biology and Biophysics
2009
University of Vermont
2006
Middlebury College
2003-2005
University of Utah
2003
Bioorthogonal ligation methods with improved reaction rates and less obtrusive components are needed for site-specifically labeling proteins without catalysts. Currently no general method exists in vivo site-specific of that combines fast rate stable, nontoxic, chemoselective reagents. To overcome these limitations, we have developed a tetrazine-containing amino acid, 1, is stable inside living cells. We genetically encoded this unique acid response to an amber codon allowing single 1 be...
14-3-3 proteins are dimeric hubs that bind hundreds of phosphorylated "clients" to regulate their function. Installing stable, functional mimics amino acids into offers a powerful strategy study function in cellular-like environments, but previous genetic code expansion (GCE) system translationally install nonhydrolyzable phosphoserine (nhpSer), with the γ-oxygen replaced CH2, site-specifically has seen limited usage. Here, we achieve 40-fold improvement this by engineering Escherichia coli...
Abstract The aromatic side-chains of phenylalanine, tyrosine, and tryptophan interact with their environments via both hydrophobic electrostatic interactions. Determining the extent to which these contribute protein function stability is not possible conventional mutagenesis. Serial fluorination a given validated method in vitro silico specifically alter characteristics, but this approach restricted select few experimental systems. Here, we report group pyrrolysine-based aminoacyl-tRNA...
ABSTRACT We previously identified a second-messenger-regulated signaling system in the environmental bacterium Pseudomonas fluorescens which controls biofilm formation response to levels of inorganic phosphate. This contains transmembrane cyclic di-GMP (c-di-GMP) receptor LapD and periplasmic protease LapG. regulates LapG ability this process large cell surface adhesin protein, LapA. While LapDG orthologs can be diverse bacteria, predictions substrates are sparse. Notably, opportunistic...
Genetic code expansion has provided the ability to site-specifically incorporate a multitude of noncanonical amino acids (ncAAs) into proteins for wide variety applications, but low ncAA incorporation efficiency can hamper utility this powerful technology. When investigating containing post-translational modification 3-nitro-tyrosine (nitroTyr), we developed second-generation amino-acyl tRNA synthetases (RS) that nitroTyr at efficiencies roughly an order magnitude greater than those...
The ability to site-specifically modify proteins at multiple sites in vivo will enable the study of protein function its native environment with unprecedented levels detail. Here, we present a versatile two-step strategy meet this goal involving site-specific encoding two distinct noncanonical amino acids bearing bioorthogonal handles into followed by mutually orthogonal labeling. This general approach, that call dual and labeling (DEAL), allowed us efficiently encode tetrazine-...
Stable surface adhesion of cells is one the early pivotal steps in bacterial biofilm formation, a prevalent adaptation strategy response to changing environments. In Pseudomonas fluorescens, this process regulated by Lap system and second messenger cyclic-di-GMP. High cytoplasmic levels cyclic-di-GMP activate transmembrane receptor LapD that turn recruits periplasmic protease LapG, preventing it from cleaving cell surface-bound adhesin, thereby promoting adhesion. study, we elucidate...
Genetic Code Expansion (GCE) can use TAG stop codons to guide site-specific incorporation of phosphoserine (pSer) into proteins. To eliminate prematurely truncated peptides, improve yields, and enhance the production multiphosphorylated proteins, Release Factor 1 (RF1)-deficient expression hosts were developed, yet these grew slowly their was associated with extensive misincorporation natural amino acids instead pSer. Here, we merge a healthy RF1-deficient E. coli cell line high-efficiency...
The dinucleotide second messenger c-di-GMP has emerged as a central regulator of reversible cell attachment during bacterial biofilm formation. A prominent adhesion mechanism first identified in pseudomonads combines two c-di-GMP-mediated processes: transcription large adhesin and its surface display via posttranslational proteolytic control. Here, we characterize an orthologous effector system show that it is operational Vibrio cholerae, where regulates distinct classes adhesins. Through...
Abstract The site‐specific incorporation of non‐canonical amino acids (ncAAs) into proteins is an important tool for understanding biological function. Traditionally, each new ncAA targeted requires a resource‐consuming process generating aminoacyl tRNA synthetase/tRNA CUA pairs. However, the discovery that some synthetases are “permissive”, in they can incorporate multiple ncAAs, means it no longer always necessary to develop synthetase newly desired ncAA. Developing better what factors...
Tyrosine nitration has served as a major biomarker for oxidative stress and is present in high abundance over 50 disease pathologies humans. While data mounts on specific pathways from sites of tyrosine nitration, the role these modifications still largely unclear. Strategies installing site-specific target proteins eukaryotic cells, through routes not dependent stress, would provide powerful method to address consequences nitration. Developed here Methanosarcina barkeri aminoacyl-tRNA...
Soluble butane monooxygenase (sBMO), a three-component di-iron complex expressed by the C 2 –C 9 alkane-utilizing bacterium Thauera butanivorans , was kinetically characterized measuring substrate specificities for 1 5 alkanes and product inhibition profiles. sBMO has high sequence homology with soluble methane (sMMO) shares similar range, including gaseous liquid alkanes, aromatics, alkenes halogenated xenobiotics. Results indicated that preferred (defined k cat : K m ratios). Relative...
Genetic code expansion (GCE) technologies commonly use the pyrrolysyl-tRNA synthetase (PylRS)/tRNA