Sulfo-SMCC Prevents Annealing of Taxol-Stabilized Microtubules In Vitro

Microtubule structure and functions have been widely studied in vitro and in cells. Research has shown that cysteines on tubulin play a crucial role in the polymerization of microtubules. Here, we show that blocking sulfhydryl groups of cysteines in taxol-stabilized polymerized microtubules with a commonly used chemical crosslinker prevents temporal end-to-end annealing of microtubules in vitro. This can dramatically affect the length distribution of the microtubules. The crosslinker sulfosuccinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate, sulfo-SMCC, consists of a maleimide and an N-hydroxysuccinimide ester group to bind to sulfhydryl groups and primary amines, respectively. Interestingly, addition of a maleimide dye alone does not show the same interference with annealing in stabilized microtubules. This study shows that the sulfhydryl groups of cysteines of tubulin that are vital for the polymerization are also important for the subsequent annealing of microtubules.Comment: 3 figure

A symmetrical method to obtain shear moduli from microrheology

Passive microrheology typically deduces shear elastic loss and storage moduli from displacement time series or mean-squared displacement (MSD) of thermally fluctuating probe particles in equilibrium materials. Common data analysis methods use either Kramers-Kronig (KK) transformations or functional fitting to calculate frequency-dependent loss and storage moduli. We propose a new analysis method for passive microrheology that avoids the limitations of both of these approaches. In this method, we determine both real and imaginary components of the complex, frequency-dependent response function $\chi(\omega) = \chi^{\prime}(\omega)+i\chi^{\prime\prime}(\omega)$ as direct integral transforms of the MSD of thermal particle motion. This procedure significantly improves the high-frequency fidelity of $\chi(\omega)$ relative to the use of KK transformation, which has been shown to lead to artifacts in $\chi^{\prime}(\omega)$. We test our method on both model data and experimental data. Experiments were performed on solutions of worm-like micelles and dilute collagen solutions. While the present method agrees well with established KK-based methods at low frequencies, we demonstrate significant improvement at high frequencies using our symmetric analysis method, up to almost the fundamental Nyquist limit.Comment: 8 pages, 4 figure

Topology counts: force distributions in circular spring networks

Filamentous polymer networks govern the mechanical properties of many biological materials. Force distributions within these networks are typically highly inhomogeneous and, although the importance of force distributions for structural properties is well recognized, they are far from being understood quantitatively. Using a combination of probabilistic and graph-theoretical techniques we derive force distributions in a model system consisting of ensembles of random linear spring networks on a circle. We show that characteristic quantities, such as mean and variance of the force supported by individual springs, can be derived explicitly in terms of only two parameters: (i) average connectivity and (ii) number of nodes. Our analysis shows that a classical mean-field approach fails to capture these characteristic quantities correctly. In contrast, we demonstrate that network topology is a crucial determinant of force distributions in an elastic spring network.Comment: 5 pages, 4 figures. Missing labels in Fig. 4 added. Reference fixe

Robust long-distance entanglement and a loophole-free Bell test with ions and photons

Two trapped ions that are kilometers apart can be entangled by the joint detection of two photons, each coming from one of the ions, in a basis of entangled states. Such a detection is possible with linear optical elements. The use of two-photon interference allows entanglement distribution without interferometric sensitivity to the path length of the photons. The present method of creating entangled ions also opens up the possibility of a loophole-free test of Bell's inequalities.Comment: published versio

Radiative Inflation and Dark Energy

We propose a model based on radiative symmetry breaking that combines inflation with Dark Energy and is consistent with the WMAP 7-year regions. The radiative inflationary potential leads to the prediction of a spectral index 0.955 \lesssim n_S \lesssim 0.967 and a tensor to scalar ratio 0.142 \lesssim r \lesssim 0.186, both consistent with current data but testable by the Planck experiment. The radiative symmetry breaking close to the Planck scale gives rise to a pseudo Nambu-Goldstone boson with a gravitationally suppressed mass which can naturally play the role of a quintessence field responsible for Dark Energy. Finally, we present a possible extra dimensional scenario in which our model could be realised.Comment: 15 pages, 4 figures; v2: references added, appendix added, Section 5 slightly modified; content matches published versio

Swelling and Softening of the Cowpea Chlorotic Mottle Virus in Response to pH Shifts

AbstractCowpea chlorotic mottle virus (CCMV) forms highly elastic icosahedral protein capsids that undergo a characteristic swelling transition when the pH is raised from 5 to 7. Here, we performed nano-indentation experiments using an atomic force microscope to track capsid swelling and measure the shells’ Young’s modulus at the same time. When we chelated Ca2+ ions and raised the pH, we observed a gradual swelling of the RNA-filled capsids accompanied by a softening of the shell. Control experiments with empty wild-type virus and a salt-stable mutant revealed that the softening was not strictly coupled to the swelling of the protein shells. Our data suggest that a pH increase and Ca2+ chelation lead primarily to a loosening of contacts within the protein shell, resulting in a softening of the capsid. This appears to render the shell metastable and make swelling possible when repulsive forces among the capsid proteins become large enough, which is known to be followed by capsid disassembly at even higher pH. Thus, softening and swelling are likely to play a role during inoculation

Combined macro- and micro-rheometer for use with Langmuir monolayers

A Langmuir monolayer trough that is equipped for simultaneous microrheology and standard rheology measurements has been constructed. The central elements are the trough itself with a full range of optical tools accessing the air-water interface from below the trough and a portable knife-edge torsion pendulum that can access the interface from above. The ability to simultaneously measure the mechanical response of Langmuir monolayers on very different lengths scales is an important step in for our understanding of the mechanical response of such systems