Functional split and crosslinking of the membrane domain of the β subunit of proton-translocating transhydrogenase from Escherichia coli

Proton pumping nicotinamide nucleotide transhydrogenase from Escherichia coli contains an α subunit with the NAD(H)-binding domain I and a β subunit with the NADP(H)-binding domain III. The membrane domain (domain II) harbors the proton channel and is made up of the hydrophobic parts of the α and β subunits. The interface in domain II between the α and the β subunits has previously been investigated by cross-linking loops connecting the four transmembrane helices in the α subunit and loops connecting the nine transmembrane helices in the β subunit. However, to investigate the organization of the nine transmembrane helices in the β subunit, a split was introduced by creating a stop codon in the loop connecting transmembrane helices 9 and 10 by a single mutagenesis step, utilizing an existing downstream start codon. The resulting enzyme was composed of the wild-type α subunit and the two new peptides β1 and β2. As compared to other split membrane proteins, the new transhydrogenase was remarkably active and catalyzed activities for the reduction of 3-acetylpyridine-NAD + by NADPH, the cyclic reduction of 3-acetylpyridine-NAD + by NADH (mediated by bound NADP(H)), and proton pumping, amounting to about 50-107% of the corresponding wild-type activities. These high activities suggest that the α subunit was normally folded, followed by a concerted folding of β1 + β2. Cross-linking of a βS105C-βS237C double cysteine mutant in the functional split cysteine-free background, followed by SDS-PAGE analysis, showed that helices 9, 13, and 14 were in close proximity. This is the first time that cross-linking between helices in the same β subunit has been demonstrated.

All-optical regenerative memory using a single device

In recent years the optical domain has been traditionally reserved for node-to-node transmission with the processing and switching achieved entirely in the electrical domain. However, with the constantly increasing demand for bandwidth and the resultant increase in transmission speeds, there is a very real fear that current electronic technology as used for processing will not be able to cope with future demands. Fuelled by this requirement for faster processing speeds, considerable research is currently being carried out into the potential of All-optical processing. One of the fundamental obstacles in realising All-optical processing is the requirement for All-optical buffering. Without all-optical buffers it is extremely difficult to resolve situations such as contention and congestion. Many devices have been proposed to solve this problem however none of them provide the perfect solution. The subject of this research is to experimentally demonstrate a novel all-optical memory device. Unlike many previously demonstrated optical storage devices the device under consideration utilises only a single loop mirror and a single SOA as its switch, whilst providing full regenerative capabilities required for long-term storage. I will explain some of the principles and characteristics of the device, which will then be experimentally demonstrated. The device configuration will then be studied and investigated as to its suitability for Hybrid Integrated Technology.

Processing and rendering of Fourier domain optical coherence tomography images at a line rate over 524 kHz using a graphics processing unit

In Fourier domain optical coherence tomography (FD-OCT), a large amount of interference data needs to be resampled from the wavelength domain to the wavenumber domain prior to Fourier transformation. We present an approach to optimize this data processing, using a graphics processing unit (GPU) and parallel processing algorithms. We demonstrate an increased processing and rendering rate over that previously reported by using GPU paged memory to render data in the GPU rather than copying back to the CPU. This avoids unnecessary and slow data transfer, enabling a processing and display rate of well over 524,000 A-scan/s for a single frame. To the best of our knowledge this is the fastest processing demonstrated to date and the first time that FD-OCT processing and rendering has been demonstrated entirely on a GPU.

Demonstrating doubly-differential quadrature phase shift keying in the optical domain

We report for the first time the experimental demonstration of doubly differential quadrature phase shift keying (DDQPSK) using optical coherent detection. This method is more robust against high frequency offsets (FO) than conventional single differential quadrature phase shift keying (SDQPSK) with offset compensation. DDQPSK is shown to be able to compensate large FOs (up to the baud rate) and has lower computational requirements than other FO compensation methods. DDQPSK is a simple algorithm to implement in a real-time decoder for optical burst switched network scenarios. Simulation results are also provided, which show good agreement with the experimental results for both SDQPSK and DDQPSK transmissions. © 1989-2012 IEEE.