A5072856130- Enzyme Structure and Function
- Yersinia bacterium, plague, ectoparasites research
- Bacterial Genetics and Biotechnology
- RNA and protein synthesis mechanisms
- Bacteriophages and microbial interactions
- Pharmacological Effects of Natural Compounds
- Protein purification and stability
- Protein Tyrosine Phosphatases
- Monoclonal and Polyclonal Antibodies Research
- Plant-based Medicinal Research
- Biochemical and Molecular Research
- Viral Infectious Diseases and Gene Expression in Insects
- Organometallic Complex Synthesis and Catalysis
- Asymmetric Hydrogenation and Catalysis
- Genetics, Bioinformatics, and Biomedical Research
- Protein Structure and Dynamics
- Bacillus and Francisella bacterial research
- ATP Synthase and ATPases Research
- Cancer-related Molecular Pathways
- DNA Repair Mechanisms
- Enzyme Production and Characterization
- Computational Drug Discovery Methods
- Vibrio bacteria research studies
- Ubiquitin and proteasome pathways
- Microtubule and mitosis dynamics
Frederick National Laboratory for Cancer Research
2012-2024
National Cancer Institute
2014-2024
Center for Cancer Research
2013-2024
Frederick Community College
2021
National Institutes of Health
2005-2020
University of Konstanz
2020
New York University
2020
Institute of Crystallography
2017
Fundación A LA PAR
2016
Loro Parque Fundación
2016
Abstract Although it is usually possible to achieve a favorable yield of recombinant protein in Escherichia coli , obtaining the soluble, biologically active form continues be major challenge. Sometimes this problem can overcome by fusing an aggregation‐prone polypeptide highly soluble partner. To study phenomenon greater detail, we compared ability three fusion partners—maltose‐binding (MBP), glutathione S‐transferase (GST), and thioredoxin (TRX)—to inhibit aggregation six diverse proteins...
Because of its stringent sequence specificity, the 3C-type protease from tobacco etch virus (TEV) is frequently used to remove affinity tags recombinant proteins. It unclear, however, exactly how TEV recognizes substrates with such high selectivity. The crystal structures two mutants, inactive C151A and autolysis-resistant S219D, have now been solved at 2.2- 1.8-Å resolution as complexes a substrate product peptide, respectively. enzyme does not appear perturbed by mutations in either...
Escherichia coli maltose binding protein (MBP) is commonly used to promote the solubility of its fusion partners. To investigate mechanism enhancement by MBP, we compared properties MBP proteins refolded in vitro with those corresponding purified under native conditions. We fused five aggregation-prone passenger 3 different N-terminal tags: His₆-MBP, His₆-GST and His₆. After purifying 15 denaturing conditions refolding them rapid dilution, recovered far more soluble than their GST- or...
Middle East respiratory syndrome coronavirus (MERS-CoV) is a highly pathogenic virus that causes severe illness accompanied by multi-organ dysfunction, resulting in case fatality rate of approximately 40%. As found other coronaviruses, the majority positive-stranded RNA MERS-CoV genome translated into two polyproteins, one created ribosomal frameshift, are cleaved at three sites papain-like protease and 11 3C-like (3 CL(pro)). Since 3 CL(pro) essential for viral replication, it leading...
Abstract Many proteins that accumulate in the form of insoluble aggregates when they are overproduced Escherichia coli can be rendered soluble by fusing them to E. maltose binding protein (MBP), and this will often enable fold their biologically active conformations. Yet, although it is an excellent solubility enhancer, MBP not a particularly good affinity tag for purification. To compensate shortcoming, we have engineered successfully tested Gateway destination vectors production dual His 6...
Summary RNA interference is mediated by small interfering RNAs produced members of the ribonuclease III (RNase III) family represented bacterial RNase and eukaryotic Rnt1p, Drosha Dicer. For mechanistic studies, has been a valuable model system for family. Previously, we have shown that uses two catalytic sites to create 2‐nucleotide (nt) 3′ overhangs in its products. Here, present three crystal structures complex with double‐stranded RNA, demonstrating how Mg 2+ essential formation...
Escherichia coli maltose‐binding protein (MBP) is frequently used as an affinity tag to facilitate the purification of recombinant proteins. An important additional attribute MBP its remarkable ability enhance solubility fusion partners. MBPs are present in a wide variety microorganisms including both mesophilic and thermophilic bacteria archaea. In study, we compared from six diverse ( E. , Pyrococcus furiosus Thermococcus litoralis Vibrio cholerae Thermotoga maritima Yersinia pestis )...
ADVERTISEMENT RETURN TO ISSUEPREVArticleNEXTSolution Structure of the Ras-Binding Domain c-Raf-1 and Identification Its Ras Interaction SurfaceS. Donald Emerson, Vincent S. Madison, Robert E. Palermo, David Waugh, Julie Scheffler, Kwei-Lan Tsao, Susan Kiefer, Shuang P. Liu, C. FryCite this: Biochemistry 1995, 34, 21, 6911–6918Publication Date (Print):May 30, 1995Publication History Published online1 May 2002Published inissue 30...