A5038302765- Protein Structure and Dynamics
- Hemoglobin structure and function
- Bacterial Genetics and Biotechnology
- Enzyme Structure and Function
- RNA and protein synthesis mechanisms
- Protein Interaction Studies and Fluorescence Analysis
- Mitochondrial Function and Pathology
- Innovative Microfluidic and Catalytic Techniques Innovation
- RNA modifications and cancer
- Biochemical and Structural Characterization
- Hedgehog Signaling Pathway Studies
- Mass Spectrometry Techniques and Applications
- Bacteriophages and microbial interactions
- Metabolomics and Mass Spectrometry Studies
- Genetic and Kidney Cyst Diseases
- Antimicrobial Resistance in Staphylococcus
- Metabolism and Genetic Disorders
- Epigenetics and DNA Methylation
- Cancer, Hypoxia, and Metabolism
- Cellular Automata and Applications
- Microfluidic and Capillary Electrophoresis Applications
- DNA and Nucleic Acid Chemistry
Fox Chase Cancer Center
2018-2023
Temple University Health System
2023
Nagoya University
2010-2016
The metabolite acetyl-CoA is necessary for both lipid synthesis in the cytosol and histone acetylation nucleus. two canonical precursors to nuclear-cytoplasmic compartment are citrate acetate, which processed by ATP-citrate lyase (ACLY) acyl-CoA synthetase short-chain 2 (ACSS2), respectively. It unclear whether other substantial routes nuclear-cytosolic exist. To investigate this, we generated cancer cell lines lacking ACLY ACSS2 [double knockout (DKO) cells]. Using stable isotope tracing,...
The development of tertiary structure during folding staphylococcal nuclease (SNase) was studied by time-resolved fluorescence resonance energy transfer measured using continuous- and stopped-flow techniques. Variants this two-domain protein containing intradomain interdomain donor/acceptor pairs (Trp Cys-linked fluorophore or quencher) were prepared to probe the structural evolution accompanying SNase folding. intra-domain are within β-barrel domain (Trp27/Cys64 Trp27/Cys97) pair is between...
ABSTRACT Acetyl-CoA is a central metabolite used for lipid synthesis in the cytosol and histone acetylation nucleus, among other pathways. The two major precursors to acetyl-CoA nuclear-cytoplasmic compartment are citrate acetate, which processed by ATP-citrate lyase (ACLY) acyl-CoA synthetase short-chain 2 (ACSS2), respectively. While some evidence has suggested existence of additional routes nuclear-cytosolic acetyl-CoA, such pathways remain poorly defined. To investigate this, we...
Several proteins have been shown to undergo a shift in the mechanism of ligand binding-induced folding from conformational selection (CS; precedes binding) induced fit (IF; binding folding) with increasing concentration. In previous studies coupled folding/binding reaction staphylococcal nuclease (SNase) presence substrate analogue, adenosine-3′,5′-diphosphate (prAp), we found that two phosphate groups make important energetic contributions toward stabilizing its complex native protein as...
Under mildly acidic conditions (pH 4–4.5) apomyoglobin (apoMb) adopts a partially structured equilibrium state (M-state) that structurally resembles kinetic intermediate encountered at late stage of folding to the native structure neutral pH. We have previously reported M-state is formed rapidly (<1 ms) via multistate process and thus offers unique opportunity for exploring early stages by both experimental computational techniques. In order gain structural insight into intermediates...
Protein conformational changes associated with ligand binding, especially those involving intrinsically disordered proteins, are mediated by tightly coupled intra- and intermolecular events. Such reactions often discussed in terms of two limiting kinetic mechanisms, selection (CS), where folding precedes induced fit (IF), binding folding. It has been shown that folding/binding can proceed along both CS IF pathways the flux ratio depending on conditions such as concentration. However,...
In this study, the equivalence of kinetic mechanisms formation urea-induced folding intermediates and non-native equilibrium states was investigated in apomyoglobin. Despite having similar structural properties, accumulate under different conditions via mechanisms, it remains unknown whether their involves shared or distinct mechanisms. To investigate potential formation, refolding unfolding kinetics horse apomyoglobin were measured by continuous- stopped-flow fluorescence over a time range...
Despite the major role of pH in protein folding and stability, a quantitative understanding pH-induced mechanism remains elusive. Two conventional models, Monod-Wyman-Changeux Linderstrøm-Lang smeared charge respectively, have been used to analyze formation/disruption specific native structures fluctuating non-native states. However, there are only few models that can represent overall kinetic events folding/unfolding independent properties relevant molecular species, which has hampered...
Many important biological processes such as protein folding and ligand binding are too fast to be fully resolved using conventional stopped-flow techniques. Although advances in mixer design detection methods have provided access the microsecond time regime, there is room for improvement terms of temporal resolution sensitivity. To address this need, we developed a continuous-flow mixing instrument with dead 12 27 µs (depending on solution viscosity) enhanced sensitivity, sufficient...
Continuous-flow FRET is a powerful technique to obtain site-specific information on tertiary structure formation during early stages of protein folding.We studied the chain condensation within beta-barrel domain and between alpha-helical domains SNase folding.Variants with single pair in showed an increase efficiency sub-millisecond time scale.In contrast, variant donor acceptor only after 100 ms folding.These results indicate that betabarrel adopts compact folding while forms final stages.
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directed mutation at the barster interface to demonstrate role ofthe hydrogen bonds mutated residue duTing complex formation.