# positions[].organizations[].parent[].label
College of Agriculture and Life Sciences

# overview
Our laboratory is interested in the mechanisms by which enzymes are regulated in the cell. In particular, we are interested in allosteric regulation of enzyme activity. Consequently, we are interested in understanding the nature of the conformational change in proteins that can be effected by the binding of ligands, and specifically how these changes alter the catalytic behavior of enzymes subject to allosteric regulation. We endeavor to investigate properties that are complementary to those determined by x-ray crystallography in order to develop a comprehensive picture of the structure-function relationships involved in the regulatory phenomenon. For example, we are interested in how the dynamics of protein structure might dictate the nature of an allosteric effect. Techniques and approaches that we use in the laboratory include analysis of enzyme kinetics; analysis of the thermodynamics of enzyme-ligand interactions; time-resolved and steady-state fluorescence spectroscopy; analysis of the effects of temperature and hydrostatic pressure (up to 4 kbar) on enzyme properties, site-specific mutagenesis, isothermal titration calorimetry, and molecular graphics.

# educationAndTraining[].organization.label
University of Wisconsin - Madison - (Madison, Wisconsin, United States)

# educationAndTraining[].organization.label
University of Illinois at Urbana Champaign - (Urbana, Illinois, United States)

# publications[].label
Distinguishing Binding from Allosteric Action in Escherichia Coli Phos-phofructokinase

# publications[].label
Contribution of single tryptophan residues to the fluorescence and stability of ribonuclease Sa.

# publications[].label
Fructose-6-phosphate binding to E-coli phosphofructokinase-1 measured by isothermal titration calorimetry resolves separate binding events

# publications[].label
The N-terminus of glycogen phosphorylase b is not required for activation by adenosine 5'-monophosphate.

# publications[].label
Identifying Unique Conformations in Thermus Thermophilus Phosphofructokinase using Fluorescence Phasors

# publications[].label
Overexpression and characterization of the human mitochondrial and cytosolic branched-chain aminotransferases.

# publications[].label
Branched chain aminotransferase isoenzymes. Purification and characterization of the rat brain isoenzyme.

# publications[].label
The binding of MgADP to the E187A mutant of E-coli phosphofructokinase.

# publications[].label
Propagation of the Allosteric Signal in Phosphofructokinase from Bacillus stearothermophilus Examined by Methyl-Transverse Relaxation-Optimized Spectroscopy Nuclear Magnetic Resonance.

# publications[].label
Secretory proteins move through the endoplasmic reticulum membrane via an aqueous, gated pore.

# publications[].label
Redefining the Role of the Quaternary Shift in the Allosteric Inhibition of Bacillus Stearothermophilus Phosphofructokinase

# publications[].label
Development of a pressure-resistant buffering system

# publications[].label
Isolation of a single activating allosteric interaction in phosphofructokinase from Escherichia coli.

# publications[].label
Redefining the role of the quaternary shift in Bacillus stearothermophilus phosphofructokinase.

# publications[].label
MgATP-dependent activation by phosphoenolpyruvate of the E187A mutant of Escherichia coli phosphofructokinase.

# publications[].label
The Effect of Urea on the Activity and Allosteric Coupling in Phosphofructokinase from Bacillus Stearothermophilus and Escherichia Coli

# publications[].label
Disentangling the web of allosteric communication in a homotetramer: heterotropic inhibition of phosphofructokinase from Bacillus stearothermophilus.

# publications[].label
Isolation of single heterotropic interactions in phosphofructokinase from B. stearothermophilus

# publications[].label
Evaluation of first-order kinetic transients encountered during enzyme assay.

# publications[].label
In vivo formation of hybrid tetramers of E. coli phosphofructokinase

# publications[].label
Kinetic Characterization of Human Liver Phosphofructokinase

# publications[].label
Stabilization of E. coli phosphofructokinase via newly introduced intersubunit disulfide bonds

# publications[].label
Structure of the apo form of Bacillus stearothermophilus phosphofructokinase.

# publications[].label
Contributions of multiple allosteric ligands are unique and additive

# publications[].label
Resolving the fluorescence response of E. coli carbamoyl phosphate synthetase

# publications[].label
Comparison of the inhibition by phospho(enol)pyruvate and phosphoglycolate of phosphofructokinase from B. stearothermophilus.

# publications[].label
The determination of thermodynamic allosteric parameters of an enzyme undergoing steady-state turnover.

# publications[].label
Tryptophan fluorescence reveals the presence of long-range interactions in the denatured state of ribonuclease Sa.

# publications[].label
Application of time-resolved fluorometry to the resolution of porphyrin-photoproduct mixtures.

# publications[].label
Tryptophan-shifted mutants as probes of the altered allosteric regulation exhibited by the E187a mutant of E-coli phosphofructokinase

# publications[].label
The isolation of large quantities of undamaged cellular organelles and cytosolic enzymes using a low-shear continuous tissue homogenizer.

# publications[].label
Identification of a residue important for heterotropic interactions in phosphofructokinase from Escherichia coli

# publications[].label
Persistent binding of MgADP to the E187A mutant of Escherichia coli phosphofructokinase in the absence of allosteric effects.

# publications[].label
Isolation of an individual allosteric interaction in tetrameric phosphofructokinase from Bacillus stearothermophilus.

# publications[].label
Do allosteric perturbations alter specificity?

# publications[].label
Allosteric effects of carbamoyl phosphate synthetase from Escherichia coli are entropy-driven.

# publications[].label
Quantitative analysis and interpretation of allosteric behavior.

# publications[].label
Disentangling the web of allosteric communication in a homotetramer: heterotropic inhibition in phosphofructokinase from Escherichia coli.

# publications[].label
Quantifying the allosteric properties of Escherichia coli carbamyl phosphate synthetase: determination of thermodynamic linked-function parameters in an ordered kinetic mechanism.

# publications[].label
Glu161 and Arg162 are not required for the inhibition by phospho(enol)pyruvate of phosphofructokinase from B-stearothermophilus

# publications[].label
Differential effects of an enthalpy-affecting mutation upon single energetic interactions of E. coli phosphofructokinase

# publications[].label
The Isolated Large Subunit of E. coli Carbamoylphosphate Synthetase Deviates from Kinetic Mechanism and Allosteric Behavior of the Holoenzyme

# publications[].label
Relationship between fructose 2,6-bisphosphate activation and MgATP inhibition of rat liver phosphofructokinase at high pH. Kinetic evidence for individual binding sites linked by finite couplings.

# publications[].label
Linked-function origins of cooperativity in a symmetrical dimer.

# publications[].label
Molecular traffic lights: Examination of enzyme subunit associations using 2-photon, dual-channel fluorescence correlation spectroscopy

# publications[].label
Influence of polyethylene glycols on the kinetics of rat liver phosphofructokinase.

# publications[].label
Effect of transforming growth factor beta (TGF-beta) on ATP citrate lyase in isolated hepatocytes.

# publications[].label
Characterization of the effects on ovalbumin mRNA of aminomethyl-trimethylpsoralen photoreaction with hen oviduct mRNA.

# publications[].label
Detection of a transient interaction between sequential metabolic enzymes using 2-photon, dual-channel FCS.

# publications[].label
Comparing the effect of selective perturbation on individual allosteric interactions within E. coli and B. stearothermophilus phosphofructokinase

# publications[].label
Allosteric dominance in carbamoyl phosphate synthetase.

# publications[].label
Quantification of liver and kidney phosphofructokinase by radioimmunoassay in fed, starved and alloxan-diabetic rats.

# publications[].label
Disentangling the web of allosteric interactions in an oligomer

# publications[].label
Failure of a two-state model to describe the influence of phospho(enol)pyruvate on phosphofructokinase from Escherichia coli.

# publications[].label
ATP-dependent activation by phospho(enol)pyruvate of the E187A mutant of E-coli phosphofructokinase

# publications[].label
Impact of hapten presentation on antibody binding at lipid membrane interfaces.

# publications[].label
Quantifying the allosteric properties of Escherichia coli carbamyl phosphate synthetase: Determination of thermodynamic linked-function parameters in an ordered kinetic mechanism

# publications[].label
Baroresistant buffer mixtures for biochemical analyses.

# publications[].label
Illuminating Dynamic Regions Associated with the Heterotropic Allosteric Communication In Bacillus Stearothermophilus Phosphofructokinase

# publications[].label
An in vivo approach to isolating allosteric pathways using hybrid multimeric proteins.

# publications[].label
Rat liver phosphofructokinase: use of fluorescence polarization to study aggregation at low protein concentration.

# publications[].label
Dynamic changes spanning the subunit interface of E. coli carbamoyl phosphate synthetase

# publications[].label
Pre-steady state quantification of the allosteric influence of Escherichia coli phosphofructokinase.

# publications[].label
Peptide sequence and conformation strongly influence tryptophan fluorescence.

# publications[].label
Remote Regions Involved in Phosphoenolpyruvate Binding to Lactobacillus Delbreuckii Phosphofructokinase

# publications[].label
Use of 2-Photon Fluorescence Correlation Spectroscopy to Investigate Rat Liver Phosphofructokinase Self-Association

# publications[].label
Fluorescence and kinetic study of A179W/W311F Trp-shifted mutant of E.coli phosphofructokinase

# publications[].label
Rat liver phosphofructokinase: kinetic and physiological ramifications of the aggregation behavior.

# publications[].label
Role of gabapentin and branched chain aminotransferase isoenzymes in astrocyte neurotransmitter metabolism.

# publications[].label
A Solution NMR and Crystallographic Study of the Role of the Quaternary Shift in the Allosteric Regulation of Phosphofructokinase from B. stearothermophilus

# publications[].label
Influence of MgADP on phosphofructokinase from Escherichia coli. Elucidation of coupling interactions with both substrates.

# publications[].label
Pb2+ as modulator of protein-membrane interactions.

# publications[].label
Construction of an inducible, pfkA and pfkB deficient strain of Escherichia coli for the expression and purification of phosphofructokinase from bacterial sources.

# publications[].label
Identifing Unique Conformational Forms of Phosphofructokinase Using Fluorescence Phasor Analysis

# publications[].label
Fluorescence and stability of different TRP-shift mutants of E. coli phosphofructokinase.

# publications[].label
Influence of a sulfhydryl cross-link across the allosteric-site interface of E. coli phosphofructokinase.

# publications[].label
Synthesis and spectral characterization of sulfhydryl-reactive fluorescent probes.

# publications[].label
Propogation of the Allosteric Signal in Bacillus Stearothermophilus Phosphofructokinase Examined by Methyl-TROSY NMR

# publications[].label
Influence of pH on the regulatory kinetics of rat liver phosphofructokinase: a thermodynamic linked-function analysis.

# publications[].label
Pressure dependence of enthalpy and entropy contributions to allosteric inhibition of E. coli phosphofructokinase

# publications[].label
Structural basis for the allosteric response of phosphofructokinase from Bacillus stearothermophilus

# publications[].label
Phospho(enol)pyruvate has little effect on the conformational change induced by fructose 6-phosphate in phosphofructokinase from E-coli

# publications[].label
High pressure effects on phosphofructokinase from E-coli: Time-domain and frequency-domain fluorescence studies.

# publications[].label
Illuminating the Dynamic Changes Associated with Heterotropic Allosteric Communication in Phosphofructokinase from Bacillus stearothermophilus

# publications[].label
Continuing Inspiration: Gregorio Weber's Influence on Understanding the Basis of Allosteric Regulation of Enzymes

# publications[].label
Influence of fructose 2,6-bisphosphate on the aggregation properties of rat liver phosphofructokinase.

# publications[].label
Formation of E-coli phosphofructokinase tetramers of mixed subunit composition

# publications[].label
Resolving the allosteric interactions found within a tetrameric enzyme

# publications[].label
Three regions important for allosteric inhibition in phosphofructokinase from lactobacillus delbrueckii subspecies bulgaricus

# publications[].label
Rat liver phosphofructokinase: kinetic activity under near-physiological conditions.

# publications[].label
Effect of Hydrostatic Pressure on Allosteric Couplings in a Tryptophan-Shifted Variant of Phosphofructokinase from Bacillus Stearothermophilus

# publications[].label
Distinguishing Interactions Responsible for Phospho(enol)pyruvate Binding from Interactions that Communicate Allosteric Inhibition in E. coli Phosphofructokinase

# publications[].label
Evaluation of ligand interactions of Bacillus stearothermophilus phosphofructokinase using the fluorescence of a tryptophan-shift mutant.

# publications[].label
The effect of modifying a fully conserved residue on the allosteric response of bacillus stearothermophilds phosphofructokinase

# publications[].label
Reevaluation of the accepted allosteric mechanism of phosphofructokinase from Bacillus stearothermophilus.

# publications[].publisher
Proceedings of the National Academy of Sciences

# publications[].label
Steady-state and time-resolved fluorescence of F188W and F76W TRP-shift mutants of E. coli phosphofructokinase

# publications[].label
A method for on-line background subtraction in frequency domain fluorometry.

# publications[].label
Allosteric inhibition and activation in E. coli phosphofructokinase use the same binding sites but different pathways

# publications[].label
Kinetic Characterization of Human Liver Phosphofructokinase

# publications[].label
Manipulation and isolation of hybrid tetramers of E-coli PFK of defined subunit composition

# publications[].label
Application of three-photon excitation FCS to the study of protein oligomerization.

# publications[].label
Dependence of ATP citrate lyase activity on cell density of isolated rat hepatocytes.

# publications[].label
Examination of MgATP binding in a tryptophan-shift mutant of phosphofructokinase from Bacillus stearothermophilus.

# publications[].label
Identification of substrate contact residues important for the allosteric regulation of phosphofructokinase from Eschericia coli.

# publications[].label
Inhibition of E. coli carbomyl phosphate synthetase by pyrimidine nucleotides

# publications[].label
Effect of pressure on the stability of F188W and F76W trp-shift mutants of E.coli phosphofructokinase

# publications[].label
Resolving the fluorescence response of Escherichia coli carbamoyl phosphate synthetase: mapping intra- and intersubunit conformational changes.

# publications[].label
Reexamination of the role of the N-terminus of glycogen phosphorylase

# publications[].label
The signal sequence moves through a ribosomal tunnel into a noncytoplasmic aqueous environment at the ER membrane early in translocation.

# publications[].label
Reversible ligand-induced dissociation of a tryptophan-shift mutant of phosphofructokinase from Bacillus stearothermophilus.

# publications[].label
MgATP and fructose 6-phosphate interactions with phosphofructokinase from Escherichia coli.

# publications[].label
Purification and characterization of the periplasmic domain of serine chemoreceptor

# publications[].label
Thermodynamic thermodynamic driving forces behind allosteric couplings in phosphofructokinases from thermophilic bacteria

# publications[].label
Effects of protein-ligand associations on the subunit interactions of phosphofructokinase from B. stearothermophilus.

# publications[].label
Mechanism of substrate inhibition in Escherichia coli phosphofructokinase.

# publications[].label
Disentangling the web of allosteric communication in a homotetramer: heterotropic activation in phosphofructokinase from Escherichia coli.

# publications[].label
Perturbation of the quaternary structure and allosteric behavior of rat liver phosphofructokinase by polyethylene glycol.

# publications[].label
Binding energy is not conferred by binding site residues alone; The binding site of E. coli phosphofructokinase

# publications[].label
Energetics and the allosteric mechanism of phosphofructokinase from Bacillus stearothermophilus.

# publications[].label
Allosteric regulation in phosphofructokinase from the extreme thermophile Thermus thermophilus.

# publications[].label
Kinetic and structural characterization of phosphofructokinase from Lactobacillus bulgaricus.

# publications[].label
Equilibrium binding studies of a tryptophan-shifted mutant of phosphofructokinase from Bacillus stearothermophilus.

# publications[].label
IIAGlc inhibition of glycerol kinase: a communications network tunes protein motions at the allosteric site.

# publications[].label
Investigation of dynamic properties of tryptophan-shifted mutants of B-stearothermophilus phosphofructokinase using tryptophan fluorescence

# publications[].label
Influence of fructose 2,6-bisphosphate and MgATP on rat liver phosphofructokinase at pH 7: evidence for a complex interdependence.

# publications[].label
Fluorescence spectroscopy as a probe of the effect of phosphorylation at serine 40 of tyrosine hydroxylase on the conformation of its regulatory domain.

# publications[].label
Carbamoyl phosphate synthetase: a crooked path from substrates to products.

# publications[].label
Studies on the kinetic mechanism of oxidative phosphorylation.

# publications[].label
Effects of ligands on the mobility of an active-site loop in tyrosine hydroxylase as monitored by fluorescence anisotropy.

# publications[].label
Quantification of allosteric influence of Escherichia coli phosphofructokinase by frequency domain fluorescence.

# publications[].label
Analyses of single tryptophan variants of E. coli carbamoyl phosphate synthetase.

# publications[].label
Use of 2-Photon Fluorescence Correlation Spectroscopy and Electron Microscopy to Ellucidate the Dependence of RLPFK Self Association on Ligand Concentration

# publications[].label
Isolation of individual allosteric conduits within phosphofructokinase from Bacillus stearothermophilus

# publications[].label
Influence of substrates and MgADP on the time-resolved intrinsic fluorescence of phosphofructokinase from Escherichia coli. Correlation of tryptophan dynamics to coupling entropy.

# publications[].label
Role of coupling entropy in establishing the nature and magnitude of allosteric response.

# publications[].publisher
Proceedings of the National Academy of Sciences

# publications[].label
Examination of MgATP binding in a tryptophan-shift mutant of phosphofructokinase from Bacillus stearothermophilus

# publications[].label
The effect of introducing small cavities on the allosteric inhibition of phosphofructokinase from Bacillus stearothermophilus.

# publications[].label
Ligand Induced Dynamic Perturbations Highlight Important Regions Leading to Inhibition of Escherichia Coli Phosphofructokinase by Phosphoenolpyruvate

# publications[].label
Optimization of active site mutations in hybrid tetramers with isolated heterotropic interactions in E. coli phosphofructokinase

# publications[].label
Weak Binding of Phospho(enol)pyruvate to Phosphofructokinase from Lactobacillus delbrueckii

# publications[].label
Quantification of isolated heterotropic interactions contributing to allostery in phosphofructokinase from E. coli

# publications[].label
A steady-state kinetic method for the verification of the rapid-equilibrium assumption in allosteric enzymes.

# researcherOn[].awardedBy.label
National Science Foundation - (Arlington, Virginia, United States)

# schools[]
College of Agriculture and Life Sciences

# etdChairOf[].label
Wang, Shanzhi (2011-10). Identification of Structural Changes Associated with Regulation of Tyrosine Hydroxylase. Doctoral Dissertation.

# etdChairOf[].label
Tindall, Amanda Jean (2016-04). Expression, Purification, and Kinetic Characterization of Human Liver Phosphofructokinase. Master's Thesis.

# etdChairOf[].label
Paricharttanakul, Nilubol Monique (2005-02). Pathway to allostery:  differential routes for allosteric communication in phosphofructokinase from Escherichia coli. Doctoral Dissertation.

# etdChairOf[].label
Perez, Stephanie (2012-10). Illuminating the Heterotropic Communication of the Pair-wise Interactions in Phosphofructokinase from Bacillus stearothermophilus. Doctoral Dissertation.

# etdChairOf[].label
Tian, Xinxin (2016-08). Probing the Details of the Allosteric Inhibition in Phosphofructokinase from Thermus thermophilus. Doctoral Dissertation.

# etdChairOf[].label
Fan, Lin (2006-08). The Tsr chemoreceptor/CheW/CheA ternary complex as an allosteric enzyme. Master's Thesis.

# etdChairOf[].label
Laird, Bobby Wayne (2015-05). Distinguishing Allosteric Interactions Using Thermodynamic Analysis and the First Reported Crystal Structure of Phospho(Enol)Pyruvate Bound E. coli Phosphofructokinase. Doctoral Dissertation.

# etdChairOf[].label
Quinlan, Robert Jason (2005-02). An investigation into the role of protein-ligand interactions on obligate and transient protein-protein interactions. Doctoral Dissertation.

# etdChairOf[].label
Ortigosa, Allison Dawn (2004-09). Resolution of the pair-wise allosteric interactions found in phosphofructokinase from Bacillus stearothermophilus. Doctoral Dissertation.

# etdChairOf[].label
Whitaker, Amy Michelle (2015-12). Exploration of Thermodynamic and Structural Changes Relevant to the Allosteric Inhibition in Phosphofructokinase from Bacillus Stearothermophilus. Doctoral Dissertation.

# etdChairOf[].label
Tie, Cuijuan (2008-10). Kinetics and dynamics study on the allosteric pathway of phosphofructokinase from Escherichia coli. Doctoral Dissertation.

# etdChairOf[].label
Shubina-McGresham, Maria (2012-10). Characterization of the Allosteric Properties of Thermus thermophilus Phosphofructokinase and the Sources of Strong Inhibitor Binding Affinity and Weak Inhibitory Response. Doctoral Dissertation.

# etdChairOf[].label
Panay Escobar, Aram Joel (2011-10). Mechanistic and Structural Studies of Phenylalanine Hydroxylase from Chromobacterium violaceum. Doctoral Dissertation.

# etdChairOf[].label
Henderson Pozzi, Michelle (2011-08). Flavin Amine Oxidases from the Monoamine Oxidase Structural Family Utilize a Hydride Transfer Mechanism. Doctoral Dissertation.

# etdChairOf[].label
Bigley, Andrew N. (2010-07). Investigations on the Mechanism of Allosteric Activtion of Rabbit Muscle Glycogen Phosphorylase b by AMP. Doctoral Dissertation.

# etdChairOf[].label
Mosser, Rockann Elizabeth (2011-10). A Structural and Kinetic Study into the Role of the Quaternary Shift in Bacillus stearothermophilus Phosphofructokinase. Doctoral Dissertation.

# etdChairOf[].label
Ferguson, Scarlett Blair (2012-10). Understanding Weak Binding for Phospho(enol)pyruvate to the Allosteric Site of Phosphofructokinase from Lactobacillus delbrueckii subspecies bulgaricus. Doctoral Dissertation.