Manipulation and growth of birefringent protein crystals in optical tweezers

We report on the trapping, rotation, and in-situ growth of birefringent tetragonal lysozyme crystals in optical tweezers operating at a wavelength of 1070 nm. Variation of the pH and lysozyme concentration of the solution during growth was used to alter the length to width ratio of the crystals, and hence their orientation in the tweezers. Crystals with the optical axis skewed or perpendicular to the trapping-beam axis could be rotated by changing the orientation of linearly polarized light. We observed spontaneous spinning of some asymmetric crystals in the presence of linearly polarized light, due to radiation pressure effects. Addition of protein to the solution in the tweezers permitted real-time observation of crystal growth. (C) 2004 Optical Society of America.

Cytochrome c(551) from Starkeya novella - Characterization, spectroscopic properties, and phylogeny of a diheme protein of the SoxAX family

Cytochromes from the SoxAX family have a major role in thiosulfate oxidation via the thiosulfate-oxidizing multi-enzyme system (TOMES). Previously characterized SoxAX proteins from Rhodovulum sulficlophilum and Paracoccus pantotrophus contain three heme c groups, two of which are located on the SoxA subunit. In contrast, the SoxAX protein purified from Starkeya novella was found to contain only two heme groups. Mass spectrometry showed that a disulfide bond replaced the second heme group found in the diheme SoxA subunits. Apparent molecular masses of 27,229 +/- 10.3 Da and 20,258.6 +/- 1 Da were determined for SoxA and SoxX with an overall mass of 49.7 kDa, indicating a heterodimeric structure. Optical redox potentiometry found that the two heme cofactors are reduced at similar potentials (versus NHE) that are as follows: + 133 mV (pH 6.0); + 104 mV (pH 7.0); +49 (pH 7.9) and +10 mV (pH 8.7). EPR spectroscopy revealed that both ferric heme groups are in the low spin state, and the spectra were consistent with one heme having a His/Cys axial ligation and the other having a His/Met axial ligation. The His/Cys ligated heme is present in different conformational states and gives rise to three distinct signals. Amino acid sequencing was used to unambiguously assign the protein to the encoding genes, soxAX, which are part of a complete sox gene cluster found in S. novella. Phylogenetic analysis of soxA- and soxX-related gene sequences indicates a parallel development of SoxA and SoxY, with the diheme and monoheme SoxA sequences located on clearly separated branches of a phylogenetic tree.

Transition metal complexes as mediator-titrants in protein redox potentiometry

A selection of nine macrocyclic Fe-III/II and Co-III/II transition metal complexes has been chosen to serve as a universal set of mediator-titrants in redox potentiometry of protein samples. The potential range spanned by these mediators is approximately from +300 to -700 mV vs the normal hydrogen electrode, which covers the range of most protein redox potentials accessible in aqueous solution. The complexes employed exhibit stability in both their oxidized and their reduced forms as well as pH-independent redox potentials within the range 6 < pH < 9. The mediators were also chosen on the basis of their very weak visible absorption maxima in both oxidation states, which will enable (for the first time) optical redox potentiometric titrations of proteins with relatively low extinction coefficients. This has previously been impractical with organic mediators, such as indoles, viologens and quinones, whose optical spectra interfere strongly with those of the protein.

Experiments with Bose-Einstein condensates on an atom chip

We describe the production of BECs on a new type of atom chip based on silver foil. Our atom chip is fabricated with thick wires capable of carrying currents of several amperes without overheating. The silver surface is highly reflective to light resonant with optical transitions used for Rb. The pattern on the chip consists of two parallel Z-trap wires, capable of producing two-wire guide, and two additional endcap wires for varying the axial confinement. Condensates are produced in magnetic microtraps formed within 1 mm of surface of the chip. We have observed the fragmentation of cold atom clouds when brought close to the chip surface. This results from a perturbed trapping potential caused by nanometer deviations of the current path through the wires on the chip. We present results of fragmentation of cold clouds at distances below 100 µm from the wires and investigate the origin of the deviating current. The fragmentation has different characteristics to those seen with copper conductors. The dynamics of atoms in these microtraps is also investigated. ©2005 COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Growth of crystals in optical tweezers

We report here on the use of optical tweezers in the growth and manipulation of protein and inorganic crystals. Sodium chloride and hen egg-white lysozyme crystals were grown in a batch process, and then seeds from the solution were introduced into the optical tweezers. The regular and controllable shape and the known optical birefringence in these structures allowed a detailed study of the orientation effects in the beam due to both polarization and gradient forces. Additionally, we determined that the laser tweezers could be used to suspend a crystal for three-dimensional growth under varying conditions. Studies included increasing the protein concentration, thermal cycling, and a diffusion-induced increase in precipitant concentration. Preliminary studies on the use of the tweezers to create a localized seed for growth from polyethylene oxide solutions are also reported.

A real-time asymmetric multiprocessor-reconfigurable system-on-chip architecture

We propose an asymmetric multi-processor SoC architecture, featuring a master CPU running uClinux, and multiple loosely-coupled slave CPUs running real-time threads assigned by the master CPU. Real-time SoC architectures often demand a compromise between a generic platform for different applications, and application-specific customizations to achieve performance requirements. Our proposed architecture offers a generic platform running a conventional embedded operating system providing a traditional software-oriented development approach, while multiple slave CPUs act as a dedicated independent real-time threads execution unit running in parallel of master CPU to achieve performance requirements. In this paper, the architecture is described, including the application / threading development environment. The performance of the architecture with several standard benchmark routines is also analysed.