Identification of a discrete intermediate in the assembly disassembly of physalis mottle tymovirus through mutational analysis

Assembly intermediates of icosahedral viruses are usually transient and are difficult to identify. In the present investigation, site-specific and deletion mutants of the coat protein gene of physalis mottle tymovirus (PhMV) were used to delineate the role of specific amino acid residues in the assembly of the virus and to identify intermediates in this process. N-terminal 30, 34, 35 and 39 amino acid deletion and single C-terminal (N188) deletion mutant proteins of PhMV were expressed in Escherichia coli. Site-specific mutants H69A, C75A, W96A, D144N, D144N-T151A, K143E and N188A were also constructed and expressed. The mutant protein lacking 30 amino acid residues from the N terminus self-assembled to T = 3 particles in vivo while deletions of 34, 35 and 39 amino acid residues resulted in the mutant proteins that were insoluble. Interestingly, the coat protein (pR PhCP) expressed using pRSET B vector with an additional 41 amino acid residues at the N terminus also assembled into T = 3 particles that were more compact and had a smaller diameter. These results demonstrate that the amino-terminal segment is flexible and either the deletion or addition of amino acid residues at the N terminus does not affect T = 3 capsid assembly, in contrast, the deletion of even a single residue from the C terminus (PhN188 Delta 1) resulted in capsids that were unstable. These capsids disassembled to a discrete intermediate with a sedimentation coefficent of 19.4 S. However, the replacement of C-terminal asparagine 188 by alanine led to the formation of stable capsids. The C75A and D144N mutant proteins also assembled into capsids that were as stable as the pR PhCP, suggesting that C75A and D144 are not crucial for the T = 3 capsid assembly. pR PhW96A and pR PhD144N-T151A mutant proteins failed to form capsids and were present as heterogeneous aggregates. Interestingly, the pR PhK143E mutant protein behaved in a manner similar to the C-terminal deletion protein in forming unstable capsids. The intermediate with an s value of 19.4 S was the major assembly product of pR PhH69A mutant protein and could correspond to a 30mer. It is possible that the assembly or disassembly is arrested at a similar stage in pR PhN188 Delta 1, pR PhH69A and pR PhK143E mutant proteins.

A note on synthesis of the general, two-degree-of-freedom linkage

A graphical method is presented for synthesis of the general, seven-link, two-degree-of-freedom plane linkage to generate functions of two variables. The method is based on point position reduction and permits synthesis of the linkage to satisfy upto six arbitrarily selected precision positions.

General classical theory of spinning particles in a meson field

An exact classical theory of the motion of a point dipole in a meson field is given which takes into account the effects of the reaction of the emitted meson field. The meson field is characterized by a constant $\chi =\mu /\hslash $ of the dimensions of a reciprocal length, $\mu $ being the meson mass, and as $\chi \rightarrow $ 0 the theory of this paper goes over continuously into the theory of the preceding paper for the motion of a spinning particle in a Maxwell field. The mass of the particle and the spin angular momentum are arbitrary mechanical constants. The field contributes a small finite addition to the mass, and a negative moment of inertia about an axis perpendicular to the spin axis. A cross-section (formula (88 a)) is given for the scattering of transversely polarized neutral mesons by the rotation of the spin of the neutron or proton which should be valid up to energies of 10$^{9}$ eV. For low energies E it agrees completely with the old quantum cross-section, having a dependence on energy proportional to p$^{4}$/E$^{2}$ (p being the meson momentum). At higher energies it deviates completely from the quantum cross-section, which it supersedes by taking into account the effects of radiation reaction on the rotation of the spin. The cross-section is a maximum at E $\sim $ 3$\cdot $5$\mu $, its value at this point being 3 $\times $ 10$^{-26}$ cm.$^{2}$, after which it decreases rapidly, becoming proportional to E$^{-2}$ at high energies. Thus the quantum theory of the interaction of neutrons with mesons goes wrong for E $\gtrsim $ 3$\mu $. The scattering of longitudinally polarized mesons is due to the translational but not the rotational motion of the dipole and is at least twenty thousand times smaller. With the assumption previously made by the present author that the heavy partilesc may exist in states of any integral charge, and in particular that protons of charge 2e and - e may occur in nature, the above results can be applied to charged mesons. Thus transversely polarised mesons should undergo a very big scattering and consequent absorption at energies near 3$\cdot $5$\mu $. Hence the energy spectrum of transversely polarized mesons should fall off rapidly for energies below about 3$\mu $. Scattering plays a relatively unimportant part in the absorption of longitudinally polarized mesons, and they are therefore much more penetrating. The theory does not lead to Heisenberg explosions and multiple processes.

Covalent assembly-disassembly of poly(ether imine) dendritic macromolecular monomers and megamers

Poly(ether imine) dendritic macromolecules were undertaken to study the reversible dendrimer monomer-megamer assembly-disassembly in aqueous solutions. Synthesis of thiol functionalized poly(ether imine) (PETIM) dendrimers and their covalent aggregation behavior in the aqueous solution of ethanol/water (2:1) is demonstrated. The dendritic megamers were characterized using microscopic techniques. Kinetics of the aggregation behavior was followed using turbidity measurements, light-scattering and atomic force microscopic techniques. Inherent luminescence behavior of PETIM dendrimer monomers was retained in the dendrimer megamers also, which allowed visualization of the megamers through confocal microscopy. Extent of thiol functionalities that remained after the megamer assembly was estimated through Ellman's assay. Subsequent to megamer assembly, disassembly of megamers to dendrimer monomers was conducted, using dithiothreitol reagent. Water-insoluble sudan I dye was encapsulated in dendrimer megamer and subsequent release profile was assessed during the disassembly in aqueous solutions. The studies were conducted using first, second and third generations, representing 4, 8 and 16 sulfhydryl groups at their peripheries of dendrimers, respectively. (C) 2014 Elsevier Ltd. All rights reserved.

Modulation of Electronic and Self-Assembly Properties of a Donor-Acceptor-Donor-Based Molecular Materials via Atomistic Approach

The performance of molecular materials in optoelectronic devices critically depends upon their electronic properties and solid-state structure. In this report, we have synthesized sulfur and selenium based (T4BT and T4BSe) donor-acceptor-donor (D-A-D) organic derivatives in order to understand the structure-property correlation in organic semiconductors by selectively tuning the chalcogen atom. The photophysical properties exhibit a significant alteration upon varying a single atom in the molecular structure. A joint theoretical and experimental investigation suggests that replacing sulfur with selenium significantly reduces the band gap and molar absorption coefficient because of lower electronegativity and ionization potential of selenium. Single-crystal X-ray diffraction analysis showed differences in their solid-state packing and intermolecular interactions. Subsequently, difference in the solid-state packing results variation in self-assembly. Micorstructural changes within these materials are correlated to their electrical resistance variation, investigated by conducting probe atomic force microscopy (CP-AFM) measurements. These results provide useful guidelines to understand the fundamental properties of D-A-D materials prepared by atomistic modulation.