共查询到20条相似文献,搜索用时 15 毫秒
1.
The nanomechanical properties of beta-galactan, a 1 --> 4 linked beta-d-galactose polysaccharide, were investigated with AFM-based single-molecule force spectroscopy. AFM captured a unique plateau at 640 pN in the force spectrogram of beta-galactan, which is significantly different than the plateau at 280 pN in the force spectrogram of amylose. Thus, our results demonstrate that force spectroscopy is able to discriminate between sugar isomers in which axial and equatorial bonds at C1 and C4 are swapped. 相似文献
2.
Liu C Jiang Z Zhang Y Wang Z Zhang X Feng F Wang S 《Langmuir : the ACS journal of surfaces and colloids》2007,23(18):9140-9142
In this letter, we report on the direct measurement of the intercalation interactions between acridine and double-stranded DNA (dsDNA) using single molecule force spectroscopy. The interaction between acridine and dsDNA is broken by force of 36 pN at a loading rate of 5.0 nN/s. The most probable rupture force between acridine and dsDNA is dependent on the loading rate, indicating that the binding of acridine and dsDNA is a dynamic process. The combination of SMFS experimental data with the theoretical model clearly suggests the presence of two energy barriers along with an unbinding trajectory of acridine-dsDNA. 相似文献
3.
Kamper SG Porter-Peden L Blankespoor R Sinniah K Zhou D Abell C Rayment T 《Langmuir : the ACS journal of surfaces and colloids》2007,23(25):12561-12565
In this communication, we report on the interaction landscape of an active site-specific enzyme-inhibitor complex by single-molecule force spectroscopy. Electrostatic immobilization was employed to orient a carbonic anhydrase enzyme on a positively charged surface so its active site is pointing upward. This approach to immobilization effectively increases the number of specific interactions measured between the zinc ion of the active site on carbonic anhydrase and a sulfonamide inhibitor tethered to an atomic force microscope (AFM) probe. Further, it reduces the time required for data collection and thereby minimizes the possible mechanical damage to the probe and contamination of the enzyme surface. The rupture force measured at various loading rates is interpreted in terms of a single energy barrier for the carbonic anhydrase enzyme-sulfonamide inhibitor complex from which the kinetic and thermodynamic parameters were estimated on the basis of microscopic models and were compared to the Bell-Evans model. The dissociation rate for the enzyme-inhibitor complex was found to be significantly faster (~35 times) than the natural spontaneous dissociation rate. 相似文献
4.
Gexiao Sun 《European Polymer Journal》2005,41(4):663-667
Using an atomic force microscope (AFM) the interaction between an AFM tip and a planar silicon oxide surface has been measured across poly(dimethylsiloxane) (PDMS, MW = 18 000). Due to the small radius of curvature of the AFM tip the hydrodynamic repulsion of the tip was negligible and forces could be measured in equilibrium. This is confirmed by the fact that force-versus-distance curves measured at different approaching velocities were indistinguishable. In equilibrium a repulsive force was observed which could best be described by a power law, F ∝ 1/d2.5 where d is the distance. 相似文献
5.
Single-molecule force spectroscopy has become a valuable tool for the investigation of intermolecular energy landscapes for a wide range of molecular associations. Atomic force microscopy (AFM) is often used as an experimental technique in these measurements, and the Bell-Evans model is commonly used in the statistical analysis of rupture forces. Most applications of the Bell-Evans model consider a constant loading rate of force applied to the intermolecular bond. The data analysis is often inconsistent because either the probe velocity or the apparent loading rate is being used as an independent parameter. These approaches provide different results when used in AFM-based experiments. Significant variations in results arise from the relative stiffness of the AFM force sensor in comparison with the stiffness of polymeric tethers that link the molecules under study to the solid surfaces. An analytical model presented here accounts for the systematic errors in force-spectroscopy parameters arising from the nonlinear loading induced by polymer tethers. The presented analytical model is based on the Bell-Evans model of the kinetics of forced dissociation and on the asymptotic models of tether stretching. The two most common data reduction procedures are analyzed, and analytical expressions for the systematic errors are provided. The model shows that the barrier width is underestimated and that the dissociation rate is significantly overestimated when force-spectroscopy data are analyzed without taking into account the elasticity of the polymeric tether. Systematic error estimates for asymptotic freely jointed chain and wormlike chain polymer models are given for comparison. The analytical model based on the asymptotic freely jointed chain stretching is employed to analyze and correct the results of the double-tether force-spectroscopy experiments of disjoining "hydrophobic bonds" between individual hexadecane molecules that are covalently tethered via poly(ethylene glycol) linkers of different lengths to the substrates and to the AFM probes. Application of the correction algorithm decreases the spread of the data from the mean value, which is particularly important for measurements of the dissociation rate, and increases the barrier width to 0.43 nm, which might be indicative of the theoretically predicted hydrophobic dewetting. 相似文献
6.
The mechanical properties of alkanethiol monolayers on Au(111) in KOH solution have been studied by force spectroscopy. The analysis of the vertical force versus penetration curves showed that monolayer penetration is a stepped process that combines elastic regions with sudden penetration events. The structural meaning of these events can be explained both by the creation of gauche defects on the hydrocarbon chains and by a cooperative molecular tilting model proposed by Barrena et al. [J. Chem. Phys. 113, 2413 (2000)]. The validity of these models for alkanethiol monolayers of different compactness and chain length has been discussed. The Young's modulus (E) of the monolayers has been calculated by using a recently developed model which considers the thickness of the monolayer as a parameter, thus allowing a decoupling of the mechanical properties of the thiol layer from those of the Au(111) substrate. As a result, the calculated E values are in the range of 50-150 Pa, which are remarkably lower than those previously reported in the literature. 相似文献
7.
Anselmetti D Bartels FW Becker A Decker B Eckel R McIntosh M Mattay J Plattner P Ros R Schäfer C Sewald N 《Langmuir : the ACS journal of surfaces and colloids》2008,24(4):1365-1370
Tunable and switchable interaction between molecules is a key for regulation and control of cellular processes. The translation of the underlying physicochemical principles to synthetic and switchable functional entities and molecules that can mimic the corresponding molecular functions is called reverse molecular engineering. We quantitatively investigated autoinducer-regulated DNA-protein interaction in bacterial gene regulation processes with single atomic force microscopy (AFM) molecule force spectroscopy in vitro, and developed an artificial bistable molecular host-guest system that can be controlled and regulated by external signals (UV light exposure and thermal energy). The intermolecular binding functionality (affinity) and its reproducible and reversible switching has been proven by AFM force spectroscopy at the single-molecule level. This affinity-tunable optomechanical switch will allow novel applications with respect to molecular manipulation, nanoscale rewritable molecular memories, and/or artificial ion channels, which will serve for the controlled transport and release of ions and neutral compounds in the future. 相似文献
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9.
Time and distance dependence of reversible polymer bridging followed by single-molecule force spectroscopy 总被引:1,自引:0,他引:1
Serpe MJ Rivera M Kersey FR Clark RL Craig SL 《Langmuir : the ACS journal of surfaces and colloids》2008,24(9):4738-4742
Polymer bridging between surfaces plays an important role in a range of fundamental processes in the material and life sciences. Bridges formed by main-chain reversible polymers differ from their covalent analogs in that they can dynamically adjust their size and shape in response to external stimuli and have the potential to reform following bond scission. In this work, the time and distance dependence of main-chain reversible polymer bridge formation are studied using an atomic force microscope. The bridging process was studied using single-molecule force spectroscopy, and its dependence on the distance between surfaces and equilibration time was probed. The number of bridges formed decreases as the gap width increases, from approximately 2 bridges per 14 s equilibration at separations of 5-15 nm to approximately 0.5 bridges per 14 s equilibration at separations of 35-45 nm. The kinetics of bridge formation appear to be slightly faster at smaller separations. 相似文献
10.
Tachikawa T Cui SC Fujitsuka M Majima T 《Physical chemistry chemical physics : PCCP》2012,14(12):4244-4249
Graphene oxide (GO) nanosheets have received a great deal of attention for a wide range of applications from optoelectronic devices to biological sensors. We now report a mechanistic study of the interfacial electron transfer (ET) processes between organic dye molecule, 9-phenyl-2,3,7-trihydroxy-6-fluorone (PF), and nanometre-sized GO sheets using ensemble-averaged and single-molecule spectroscopies. The ET dynamics was characterized by the direct observation of the PF radical cation during the laser flash photolysis, and its reaction rate was determined to be ~10(11) s(-1). The single-molecule fluorescence spectroscopy was utilized to clarify the heterogeneous nature of the interfacial ET within individual composites. Their fluorescence lifetimes and spectra were found to vary from composite to composite, possibly due to the different local structures and molecular interactions. The autocorrelation analysis of fluorescence intensity trajectories also revealed the temporal fluctuation of the ET reactivity. 相似文献
11.
Constructing polyproteins consisting of identical tandem repeats of proteins provides an unambiguous method of investigating the mechanical properties of proteins at the single-molecule level using force spectroscopy techniques. Here we report a maleimide-thiol coupling-based facile method of constructing polyproteins for single-molecule force spectroscopy studies on the mechanical properties of proteins. This method allows for the construction of polyproteins in an efficient fashion under room temperature. The resultant thioether bonds are resistant to reduction and make it possible to carry out single-molecule force spectroscopy studies under various redox conditions. This novel method complements existing polyprotein engineering methods and can be easily applied to a wide variety of proteins. 相似文献
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Nanopatterned cross-linked polymers are important for applications with controlled mechanical properties. Grafted linear and cross-linked polydimethylacrylamide gels on micro- and nanopatterns were created using iniferter-driven quasi-living radical polymerization combined with conventional photolithography and nanosphere lithography. Micropatterned linear polymers reproduce the expected scaling behavior at moderate grafting density. The addition of cross-linker to the polymerization solution leads to an increased tendency of early termination as determined by AFM force spectroscopy. Similarly, nanopatterned linear polymers show reduced thickness in agreement with the expected scaling relationship for nanoisland grafts that have reduced lateral confinement. The addition of cross-linker reintroduces some of the lateral confinement for the length of polymers reported here. The mechanical properties of both the micro- and nanopatterned linear as well as cross-linked polymers were analyzed using an algorithm to objectively determine the contact point in AFM force spectroscopy and two independent Hertz-based analysis approaches. The obtained Young's moduli are close to those expected for homogeneous thick polymer films and are independent of pattern size. Our results demonstrate that polymeric nanopillars with controlled elastic modulus can be fabricated using irreversible cross-linkers. They also highlight some of the factors that must be considered for successful fabrication of grafted nanopillars of defined mechanical and structural properties. 相似文献
14.
Knotted conformation is one of the most surprising topological features found in proteins, and understanding the folding mechanism of such knotted proteins remains a challenge. Here, we used optical tweezers (OT) to investigate the mechanical unfolding and folding behavior of a knotted protein Escherichia coli tRNA (guanosine-1) methyltransferase (TrmD). We found that when stretched from its N- and C-termini, TrmD can be mechanically unfolded and stretched into a tightened trefoil knot, which is composed of ca. 17 residues. Stretching of the unfolded TrmD involved a compaction process of the trefoil knot at low forces. The unfolding pathways of the TrmD were bifurcated, involving two-state and three-state pathways. Upon relaxation, the tightened trefoil knot loosened up first, leading to the expansion of the knot, and the unfolded TrmD can then fold back to its native state efficiently. By using an engineered truncation TrmD variant, we stretched TrmD along a pulling direction to allow us to mechanically unfold TrmD and untie the trefoil knot. We found that the folding of TrmD from its unfolded polypeptide without the knot is significantly slower. The knotting is the rate-limiting step of the folding of TrmD. Our results highlighted the critical importance of the knot conformation for the folding and stability of TrmD, offering a new perspective to understand the role of the trefoil knot in the biological function of TrmD.Optical tweezers are used to stretch a knotted protein along different directions to probe its unfolding–folding behaviors, and the conformational change of its knot structure. 相似文献
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16.
In single-molecule force spectroscopy, individual molecules and complexes are often stretched by pulling devices via intervening molecular handles. Accurate interpretation of measurements from such experiments in terms of the underlying energy landscape, defined by activation barriers and intrinsic rates of transition, relies on our understanding, and proper theoretical treatment, of the effects of the pulling device and handle. Here, we present a framework based on Kramers' theory that elucidates the dependence of measured rupture forces and rates on the pulling device stiffness and attributes of the handle, contour length and persistence length. We also introduce a simple analytic model that improves prediction of activation barriers and intrinsic rates for all device stiffnesses and handle properties, thus allowing for a more reliable interpretation of experiments. Our analyses also suggests intuitive ways of displaying the measured force spectra for proper prognosis of device and handle effects and provides the range of device and handle attributes over which these effects can be neglected. 相似文献
17.
Sharma S Gillespie BM Palanisamy V Gimzewski JK 《Langmuir : the ACS journal of surfaces and colloids》2011,27(23):14394-14400
Exosomes are naturally occurring nanoparticles with unique structure, surface biochemistry, and mechanical characteristics. These distinct nanometer-sized bioparticles are secreted from the surfaces of oral epithelial cells into saliva and are of interest as oral-cancer biomarkers. We use high- resolution AFM to show single-vesicle quantitative differences between exosomes derived from normal and oral cancer patient's saliva. Compared to normal exosomes (circular, 67.4 ± 2.9 nm), our findings indicate that cancer exosome populations are significantly increased in saliva and display irregular morphologies, increased vesicle size (98.3 ± 4.6 nm), and higher intervesicular aggregation. At the single-vesicle level, cancer exosomes exhibit significantly (P < 0.05) increased CD63 surface densities. To our knowledge, it represents the first report detecting single-exosome surface protein variations. Additionally, high-resolution AFM imaging of cancer saliva samples revealed discrete multivesicular bodies with intraluminal exosomes enclosed. We discuss the use of quantitative, nanoscale ultrastructural and surface biomolecular analysis of saliva exosomes at single-vesicle- and single-protein-level sensitivities as a potentially new oral cancer diagnostic. 相似文献
18.
Interaction of human serum albumin with bendroflumethiazide studied by fluorescence spectroscopy 总被引:5,自引:0,他引:5
Pang YH Yang LL Shuang SM Dong C Thompson M 《Journal of photochemistry and photobiology. B, Biology》2005,80(2):139-144
The interactions between bendroflumethiazide (BFTZ) and human serum albumin (HSA) have been studied by fluorescence spectroscopy. Binding constants for drug attachment to the various binding sites of HSA have been measured at different temperatures in physiological buffer solution. The effect of metal ions on BFTZ interaction with HSA was also investigated. The thermodynamic parameters, DeltaH and DeltaS, have been calculated to be 49.28kJmol(-1)>0, and 258.83Jmol(-1)K(-1)>0, respectively. The distance between HSA and BFTZ, r, was determined to be 1.47nm based on F?rster's non-radiative energy transfer theory. The experimental results reveal that BFTZ has a strong ability to quench the intrinsic fluorescence of HSA through a static quenching mechanism. Furthermore, the binding constants between BFTZ and HSA are remarkably independent of temperature, and decrease in the presence of various ions, usually by about 30-55%. Hydrophobic interaction occurs between BFTZ and the sub-domain II A of HSA. 相似文献
19.
Uchiyama D Oikawa H Otomo K Nango M Dewa T Fujiyoshi S Matsushita M 《Physical chemistry chemical physics : PCCP》2011,13(24):11615-11619
As a model of photosynthetic unit (PSU), self-assembled aggregates of pigment-protein complexes from photosynthetic bacteria were prepared in a lipid bilayer by reconstitution of the light-harvesting 2 (LH2) complex and light-harvesting 1-reaction center (LH1-RC) complex through detergent removal of their micelles in the presence of lipids. By performing polarization-controlled fluorescence and fluorescence-excitation spectroscopy on single aggregates at a temperature of 5 K, the composition of individual aggregates was determined and excitation energy transfer (EET) between constituent complexes was observed. LH2 and LH1-RC from a bacterium, Rhodobacter (Rb.) sphaeroides, were found to form a trimeric aggregate in which EET takes place from one LH2 to two LH1-RCs. In contrast, a heterodimer of LH2 and LH1-RC in which EET works was found to assemble from a combination of complexes of different bacterial species, that is, LH2 from Rb. sphaeroides and LH1-RC from Rhodopseudomonas (Rps.) palustris. 相似文献
20.
Zhang W Lü X Zhang W Shen J 《Langmuir : the ACS journal of surfaces and colloids》2011,27(24):15008-15015
In this article, interactions between Bacillus subtilis single-stranded DNA binding proteins (BsSSB) and single-stranded DNA (ssDNA) were systematically studied. The effect of different molar ratios between BsSSB and ssDNA on their binding modes was first investigated by electrophoretic mobility shift assays (EMSAs). It is found that a high molar ratio of BsSSB to ssDNA can produce BsSSB-ssDNA complexes formed in the mode of two proteins binding one 65-nt (nucleotide) ssDNA whereas a low molar ratio facilitates the formation of BsSSB-ssDNA complexes in the mode of one protein binding one 65-nt ssDNA. Furthermore, two binding modes are in dynamic equilibrium. The unbinding force of BsSSB-ssDNA complexes was measured quantitatively in solutions with different salt concentrations by using AFM-based single-molecule force spectroscopy (SMFS). Our results show that the unbinding force is about 10 pN higher at high salt concentration (0.5 M NaCl) than at low salt concentration (0.1 M NaCl) and the lifetime of BsSSB-ssDNA complexes at high salt concentration is twice as long as that at low salt concentration. These results indicate that more tightly packed BsSSB-ssDNA complexes can form at high salt (0.5 M NaCl) concentration. In addition, the results of EMSA show that ssDNA, which is bound to BsSSB, can dissociate from BsSSB in the presence of the cDNA strand, indicating the dynamic nature of BsSSB-ssDNA interactions. 相似文献