Abstract:
:The heterologous expression and purification of membrane proteins represent major limitations for their functional and structural analysis. Here we describe a new method of incorporation of transmembrane proteins in planar lipid bilayer starting from 1 pmol of solubilized proteins. The principle relies on the direct incorporation of solubilized proteins into a preformed planar lipid bilayer destabilized by dodecyl-beta-maltoside or dodecyl-beta-thiomaltoside, two detergents widely used in membrane biochemistry. Successful incorporations are reported at 20 degrees C and at 4 degrees C with three bacterial photosynthetic multi-subunit membrane proteins. Height measurements by atomic force microscopy (AFM) of the extramembraneous domains protruding from the bilayer demonstrate that proteins are unidirectionally incorporated within the lipid bilayer through their more hydrophobic domains. Proteins are incorporated at high density into the bilayer and on incubation diffuse and segregate into protein close-packing areas. The high protein density allows high-resolution AFM topographs to be recorded and protein subunits organization delineated. This approach provides an alternative experimental platform to the classical methods of two-dimensional crystallization of membrane proteins for the structural analysis by AFM. Furthermore, the versatility and simplicity of the method are important intrinsic properties for the conception of biosensors and nanobiomaterials involving membrane proteins.
journal_name
Biophys Jjournal_title
Biophysical journalauthors
Milhiet PE,Gubellini F,Berquand A,Dosset P,Rigaud JL,Le Grimellec C,Lévy Ddoi
10.1529/biophysj.106.087791subject
Has Abstractpub_date
2006-11-01 00:00:00pages
3268-75issue
9eissn
0006-3495issn
1542-0086pii
S0006-3495(06)72039-2journal_volume
91pub_type
杂志文章abstract::Many proteins contain flexible structures such as loops and hinged domains. A simple root mean square deviation (RMSD) alignment of two different conformations of the same protein can be skewed by the difference between the mobile regions. To overcome this problem, we have developed a novel method to overlay two prote...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1529/biophysj.105.066654
更新日期:2006-06-15 00:00:00
abstract::Numerous membrane-transport proteins are major drug targets, and therefore a key ingredient in pharmaceutical development is the availability of reliable, efficient tools for membrane transport characterization and inhibition. Here, we present the use of evanescent-wave sensing for screening of membrane-protein-mediat...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2010.03.059
更新日期:2010-07-07 00:00:00
abstract::The nuclei of epididymal sperm, isolated from C57BL/6J and CBA/J inbred mice by their resistance to trypsin digestion, retain the shape differences of the intact sperm head. Various physical characteristics of these nuclei were measured and compared. The measurement of the projected dimensions of nuclei showed that th...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(76)85730-X
更新日期:1976-07-01 00:00:00
abstract::Some plant leaves display complex, orientation-dependent, proton nuclear magnetic resonance (1H NMR) spectra. The spectral patterns vary as the angle between the leaf surface and the applied magnetic field is varied. They also vary with temperature and with the quantity of absorbed manganous ions, but they are indepen...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(85)83826-1
更新日期:1985-11-01 00:00:00
abstract::The mathematics of the binomial model for quantal neurotransmitter release is considered in general terms, to explore what information might be extractable from statistical aspects of data. For an array of N statistically independent release sites, each with a release probability p, the compound binomial always pertai...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/s0006-3495(97)78709-5
更新日期:1997-02-01 00:00:00
abstract::In rat mesenteric arteries, smooth muscle cells exhibit intercellular calcium waves in response to local phenylephrine stimulation. These waves have a velocity of approximately 20 cells/s and a range of approximately 80 cells. We analyze these waves in a theoretical model of a population of coupled smooth muscle cells...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2010.04.031
更新日期:2010-07-21 00:00:00
abstract::Phosphatidylinositol-4,5-bisphosphate (PIP2) is an important signaling lipid in eukaryotic cell plasma membranes, playing an essential role in diverse cellular processes. The headgroup of PIP2 is highly negatively charged, and this lipid displays a high critical micellar concentration compared to housekeeping phosphol...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2019.07.047
更新日期:2019-09-03 00:00:00
abstract::We have used optical tweezers to study the elasticity of red cell membranes; force was applied to a bead attached to a permeabilized spherical ghost and the force-extension relation was obtained from the response of a second bead bound at a diametrically opposite position. Interruption of the skeletal network by disso...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(99)77139-0
更新日期:1999-12-01 00:00:00
abstract::A new method, weighted-ensemble Brownian dynamics, is proposed for the simulation of protein-association reactions and other events whose frequencies of outcomes are constricted by free energy barriers. The method features a weighted ensemble of trajectories in configuration space with energy levels dictating the prop...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(96)79552-8
更新日期:1996-01-01 00:00:00
abstract::We investigate unfolding/folding force kinetics in DNA hairpins exhibiting two and three states with newly designed short dsDNA handles (29 bp) using optical tweezers. We show how the higher stiffness of the molecular setup moderately enhances the signal/noise ratio (SNR) in hopping experiments as compared to conventi...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2011.01.071
更新日期:2011-04-06 00:00:00
abstract::Cells in a cloned population of coliform bacteria exhibit a wide range of swimming behaviors--a form of non-genetic individuality. We used computer models to examine the proposition that these variations are due to differences in the number of chemotaxis signaling molecules from one cell to the next. Simulations were ...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(98)77777-X
更新日期:1998-01-01 00:00:00
abstract::Stearoylsphingomyelin (SSM) bilayers containing 0, 22, and 50 mol % cholesterol (Chol) and a pentadecanoyl-stearoylphosphatidylcholine (15SPC) bilayer containing 22 mol % Chol were molecular dynamics simulated at two temperatures (37 degrees C and 60 degrees C). 15SPC is the best PC equivalent of SSM. The Chol effect ...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1529/biophysj.106.080887
更新日期:2006-11-15 00:00:00
abstract::The pore-forming alpha 1 subunit of L-type calcium (Ca2+) channels is the molecular target of Ca2+ channel blockers such as phenylalkylamines (PAAs). Association and dissociation rates of (-)devapamil were compared for a highly PAA-sensitive L-type Ca2+ channel chimera (Lh) and various class A Ca2+ channel mutants. Th...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(97)78056-1
更新日期:1997-07-01 00:00:00
abstract::Precise information regarding the transmembrane topology of mitochondrial porin is essential for understanding the mechanisms by which this protein functions. Porin acts as a channel in the outer membrane and interacts with small solutes and proteins to regulate mitochondrial function. The acquisition of high-resoluti...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1529/biophysj.107.115196
更新日期:2008-01-15 00:00:00
abstract::Nisin is an antimicrobial peptide used as food preservative. To gain some insights into the hypothesis that its bactericidal activity is due to the perturbation of the lipid fraction of the bacterial plasmic membrane, we have investigated the effect of nisin on model phosphatidylcholine (PC) membranes. We show that ni...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(99)76936-5
更新日期:1999-08-01 00:00:00
abstract::The spermine-induced DNA condensation is a first-order phase transition. Here, we apply a novel technique fluorescence lifetime correlation spectroscopy to analyze this transition in a greater detail. We show that the method allows for the observation of the condensed and uncondensed molecules simultaneously based sol...
journal_title:Biophysical journal
pub_type: 信件
doi:10.1529/biophysj.107.122408
更新日期:2008-02-01 00:00:00
abstract::In contrast with most inhalational anesthetics, the anesthetic gases xenon (Xe) and nitrous oxide (N(2)O) act by blocking the N-methyl-d-aspartate (NMDA) receptor. Using x-ray crystallography, we examined the binding characteristics of these two gases on two soluble proteins as structural models: urate oxidase, which ...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1529/biophysj.106.093807
更新日期:2007-01-01 00:00:00
abstract::Many different cell types are able to migrate by formation of a thin actin-based cytoskeletal extension. Recently, it became evident that this extension consists of two distinct substructures, designated lamellipodium and lamellum, which differ significantly in their kinetic and kinematic properties as well as their b...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1529/biophysj.108.134817
更新日期:2008-12-15 00:00:00
abstract::The kinetics of force production in chemically skinned trabeculae from the guinea pig were studied by laser photolysis of caged ATP in the presence of Ca2+. Preincubation of the tissue during rigor with the enzyme apyrase was used to reduce the population of MgADP-bound cross-bridges (Martin and Barsotti, 1994). In un...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(94)80676-9
更新日期:1994-11-01 00:00:00
abstract::Reattachment and healing of tendon to bone poses a persistent clinical challenge and often results in poor outcomes, in part because the mechanisms that imbue the uninjured tendon-to-bone attachment with toughness are not known. One feature of typical tendon-to-bone surgical repairs is direct attachment of tendon to s...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2014.09.049
更新日期:2015-01-20 00:00:00
abstract::The stability of the 1:1 complex of sodium ion with the dianion of guanosine 5'-monophosphate has been determined by means of a potentiometric titration employing a specific ion electrode. The stability constant for the reaction Na(+) + 5'-GMP(2-) Na(5'-GMP)(-) was found to be 2.85 +/- 0.36 M(-1) at 5 degrees C and an...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/s0006-3495(82)84564-5
更新日期:1982-06-01 00:00:00
abstract::The excitation of the squid giant axon that had been perfused intra- and extracellularly with solutions containing a high concentration of glycerol could be observed below 0 degree C. The action potential could be elicited at normal strengths of electrical stimuli. The time-course of the action potential was slowed, w...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(81)84785-6
更新日期:1981-07-01 00:00:00
abstract::Unraveling the conformation of membrane-bound viral fusion peptides is essential for understanding how those peptides destabilize the bilayer topology of lipids that is important for virus-cell membrane fusion. Here, molecular dynamics (MD) simulations were performed to investigate the conformation of the 20 amino aci...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1529/biophysj.103.024562
更新日期:2004-07-01 00:00:00
abstract::Plasmodium falciparum malaria-infected red blood cells (IRBCs), or erythrocytes, avoid splenic clearance by adhering to host endothelium. Upregulation of endothelial receptors intercellular adhesion molecule-1 (ICAM-1) and cluster of differentiation 36 (CD36) are associated with severe disease pathology. Most in vitro...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2019.11.016
更新日期:2020-01-07 00:00:00
abstract::Nerve growth factor (NGF) mimics play an important role for therapies that target the receptor tyrosine kinase A (trkA). The N-terminal fragment of the NGF (N-term@NGF) was previously demonstrated to be an important determinant for affinity and specificity in the binding to trkA. Here we use a variety of computational...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1529/biophysj.106.083519
更新日期:2006-09-15 00:00:00
abstract::Molecular dynamics simulations of the tripeptide Ala-Phe-Ala-O-tert-butyl interacting with dimyristoylphosphatidylcholine lipid bilayers have been carried out. The lipid and aqueous environments of the peptide, the alkyl chain order, and the lipid and peptide dynamics have been investigated with use of density profile...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(95)79997-0
更新日期:1995-10-01 00:00:00
abstract::Formamide-induced detubulation of rat ventricular myocytes was used to investigate the functional distribution of the Na/Ca exchanger (NCX) and Na/K-ATPase between the t-tubules and external sarcolemma. Detubulation resulted in a 32% decrease in cell capacitance, whereas cell volume was unchanged. Thus, the surface-to...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(03)74758-4
更新日期:2003-11-01 00:00:00
abstract::Eukaryotic genomes are hierarchically organized into protein-DNA assemblies for compaction into the nucleus. Nucleosomes, with the (H3-H4)2 tetrasome as a likely intermediate, are highly dynamic in nature by way of several different mechanisms. We have recently shown that tetrasomes spontaneously change the direction ...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2019.07.055
更新日期:2019-12-03 00:00:00
abstract::Cryo-electron microscopy is rapidly emerging as a powerful technique to determine the structures of complex macromolecular systems elusive to other techniques. Because many of these systems are highly dynamical, characterizing their movements is also a crucial step to unravel their biological functions. To achieve thi...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/j.bpj.2018.02.028
更新日期:2018-04-10 00:00:00
abstract::We present a theory for proton diffusion through an immobilized protein membrane perfused with an electrolyte and a buffer. Using a Nernst-Planck equation for each species and assuming local charge neutrality, we obtain two coupled nonlinear diffusion equations with new diffusion coefficients dependent on the concentr...
journal_title:Biophysical journal
pub_type: 杂志文章
doi:10.1016/S0006-3495(79)85274-1
更新日期:1979-01-01 00:00:00