950 research outputs found
Ligand Binding, Protein Fluctuations, and Allosteric Free Energy
Although the importance of protein dynamics in protein function is generally
recognized, the role of protein fluctuations in allosteric effects scarcely has
been considered. To address this gap, the Kullback-Leibler divergence (Dx)
between protein conformational distributions before and after ligand binding
was proposed as a means of quantifying allosteric effects in proteins. Here,
previous applications of Dx to methods for analysis and simulation of proteins
are first reviewed, and their implications for understanding aspects of protein
function and protein evolution are discussed. Next, equations for Dx suggest
that k_{B}TDx should be interpreted as an allosteric free energy -- the free
energy associated with changing the ligand-free protein conformational
distribution to the ligand-bound conformational distribution. This
interpretation leads to a thermodynamic model of allosteric transitions that
unifies existing perspectives on the relation between ligand binding and
changes in protein conformational distributions. The definition of Dx is used
to explore some interesting mathematical relations among commonly recognized
thermodynamic and biophysical quantities, such as the total free energy change
upon ligand binding, and ligand-binding affinities for individual protein
conformations. These results represent the beginnings of a theoretical
framework for considering the full protein conformational distribution in
modeling allosteric transitions. Early applications of the framework have
produced results with implications both for methods for coarsed-grained
modeling of proteins, and for understanding the relation between ligand binding
and protein dynamics.Comment: 18 pages; 7 figures; Second International Congress of the
Biocomputing and Physics of Complex Systems Research Institute, Zaragoza,
Spain, 8-11 Feb 2006; increase breadth of review of methods for analysis of
allosteric mechanisms; Add AIP in press; fix missing kTs in equation
Quantum crystallographic charge density of urea
Standard X-ray crystallography methods use free-atom models to calculate mean
unit cell charge densities. Real molecules, however, have shared charge that is
not captured accurately using free-atom models. To address this limitation, a
charge density model of crystalline urea was calculated using high-level
quantum theory and was refined against publicly available ultra high-resolution
experimental Bragg data, including the effects of atomic displacement
parameters. The resulting quantum crystallographic model was compared to models
obtained using spherical atom or multipole methods. Despite using only the same
number of free parameters as the spherical atom model, the agreement of the
quantum model with the data is comparable to the multipole model. The static,
theoretical crystalline charge density of the quantum model is distinct from
the multipole model, indicating the quantum model provides substantially new
information. Hydrogen thermal ellipsoids in the quantum model were very similar
to those obtained using neutron crystallography, indicating that quantum
crystallography can increase the accuracy of the X-ray crystallographic atomic
displacement parameters. The results demonstrate the feasibility and benefits
of integrating fully periodic quantum charge density calculations into ultra
high-resolution X-ray crystallographic model building and refinement.Comment: 40 pages, 4 figures, 6 table
Prediction of Functional Sites in SCOP Domains using Dynamics Perturbation Analysis
Dynamics perturbation analysis (DPA) finds regions in a protein structure where proteins are "ticklish", i.e., where interactions cause a large change in protein dynamics. Previously, such regions were shown to predict the location of native binding sites in a docking test set, but the more general applicability of DPA to the prediction of functional sites in proteins was not shown. Here we describe the results of applying an accelerated algorithm, called Fast DPA, to predict functional sites in over 50,000 SCOP domains
Visfatin reduces gap junction mediated cell-to-cell communication in proximal tubule-derived epithelial cells
Background/Aims: In the current study we examined if the adipocytokine, visfatin, alters connexin-mediated intercellular communication in proximal tubule-derived epithelial cells. Methods: The effects of visfatin (10-200ng/mL) on cell viability and cytotoxicity in HK2-cells were assessed by MTT, crystal violet and lactate dehydrogenase assays. Western blot analysis was used to confirm expression of Cx26, Cx40 and Cx43. The effect of visfatin (10-200ng/mL) on TGF-β1 secretion was confirmed by ELISA, and the effects of both TGF-β1 (2-10ng/mL) and visfatin (10-200ng/mL) on connexin expression were assessed by western blot. Functional intercellular communication was determined using transfer of Lucifer Yellow and paired-whole cell patch clamp electrophysiology. Results: In low glucose (5mM), visfatin (10-200ng/mL) did not affect membrane integrity, cytotoxicity or cell viability at 48hrs, but did evoke a concentration-dependent reduction in Cx26 and Cx43 expression. The expression of Cx40 was unaffected. At 48hrs, visfatin (10-200ng/mL) increased the secretion of TGF-β1 and the visfatin-evoked changes in connexin expression were mimicked by exogenous application of the pro-fibrotic cytokine (2-10ng/ml). Visfatin reduced dye transfer between coupled cells and decreased functional conductance, with levels falling by 63% as compared to control. Although input resistance was increased following visfatin treatment by 166%, the change was not significant as compared to control. The effects of visfatin on Cx-expression and cell-coupling were blocked in the presence of a TGF-β1 specific neutralizing antibody. Conclusions: The adipocytokine visfatin selectively evoked a non-toxic reduction in connexin expression in HK2-cells. The loss in gap-junction associated proteins was mirrored by a loss in functional conductance between coupled cells. Visfatin increased TGF-β secretion and the pattern of change for connexins expression was mimicked by exogenous application of TGF-β1. The effect of visfatin on Cx-expression and dye transfer were negated in the presence of a TGF-β1 neutralising antibody. These data suggest that visfatin reduces connexin-mediated intercellular communication in proximal tubule-derived epithelial cells via a TGF-β dependent pathway.
© 2013 S. Karger AG, Base
Cooling Fermions in an Optical Lattice by Adiabatic Demagnetization
The Fermi-Hubbard model describes ultracold fermions in an optical lattice
and exhibits antiferromagnetic long-ranged order below the N\'{e}el
temperature. However, reaching this temperature in the lab has remained an
elusive goal. In other atomic systems, such as trapped ions, low temperatures
have been successfully obtained by adiabatic demagnetization, in which a strong
effective magnetic field is applied to a spin-polarized system, and the
magnetic field is adiabatically reduced to zero. Unfortunately, applying this
approach to the Fermi-Hubbard model encounters a fundamental obstacle: the
symmetry introduces many level crossings that prevent the system from
reaching the ground state, even in principle. However, by breaking the
symmetry with a spin-dependent tunneling, we show that adiabatic
demagnetization can achieve low temperature states. Using density matrix
renormalization group (DMRG) calculations in one dimension, we numerically find
that demagnetization protocols successfully reach low temperature states of a
spin-anisotropic Hubbard model, and we discuss how to optimize this protocol
for experimental viability. By subsequently ramping spin-dependent tunnelings
to spin-independent tunnelings, we expect that our protocol can be employed to
produce low-temperature states of the Fermi-Hubbard Model.Comment: References adde
Adenosine A1 receptor activation mediates the developmental shift at layer 5 pyramidal cell synapses and is a determinant of mature synaptic strength
During the first postnatal month glutamatergic synapses between layer 5 pyramidal cells in the rodent neocortex switch from an immature state exhibiting high probability of neurotransmitter release, large unitary amplitude and synaptic depression to a mature state with decreased probability of release, smaller unitary amplitude and synaptic facilitation. Using paired recordings, we demonstrate that the developmental shift in release probability at synapses between rat somatosensory layer 5 thick-tufted pyramidal cells is due to a higher and more heterogeneous activation of presynaptic adenosine A1 receptors. Immature synapses under control conditions exhibited distributions of CV, failure rate and release probability that were almost coincident with the A1 receptor blocked condition; however, mature synapses under control conditions exhibited much broader distributions that spanned those of both the A1 receptor agonised and antagonised conditions. Immature and mature synapses expressed A1 receptors with no observable difference in functional efficacy and therefore the heterogeneous A1 receptor activation seen in the mature neocortex is due to increased adenosine concentrations that vary between synapses. Given the central role demonstrated for A1 receptor activation in determining synaptic amplitude and the statistics of transmission between mature layer 5 pyramidal cells, the emplacement of adenosine sources and sinks near the synaptic terminal could constitute a novel form of long-term synaptic plasticity
Model of Transcriptional Activation by MarA in Escherichia coli
We have developed a mathematical model of transcriptional activation by MarA
in Escherichia coli, and used the model to analyze measurements of
MarA-dependent activity of the marRAB, sodA, and micF promoters in mar-rob-
cells. The model rationalizes an unexpected poor correlation between the
mid-point of in vivo promoter activity profiles and in vitro equilibrium
constants for MarA binding to promoter sequences. Analysis of the promoter
activity data using the model yielded the following predictions regarding
activation mechanisms: (1) MarA activation of the marRAB, sodA, and micF
promoters involves a net acceleration of the kinetics of transitions after RNA
polymerase binding, up to and including promoter escape and message elongation;
(2) RNA polymerase binds to these promoters with nearly unit occupancy in the
absence of MarA, making recruitment of polymerase an insignificant factor in
activation of these promoters; and (3) instead of recruitment, activation of
the micF promoter might involve a repulsion of polymerase combined with a large
acceleration of the kinetics of polymerase activity. These predictions are
consistent with published chromatin immunoprecipitation assays of interactions
between polymerase and the E. coli chromosome. A lack of recruitment in
transcriptional activation represents an exception to the textbook description
of activation of bacterial sigma-70 promoters. However, use of accelerated
polymerase kinetics instead of recruitment might confer a competitive advantage
to E. coli by decreasing latency in gene regulation.Comment: 30 pages, 2 figure
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