MATCHED ARCHITECTURES FOR SIGNAL PROCESSING AND CONTROL

Fast processing environments for real-time data acquisition, data processing and control applications may be realised using very different architectures. State of the art systems generally employ multiprocessors and parallel processing having a dedicated architecture such as systolic arrays to support computation-intensive signal processing tasks such as, for instance, convolution, filtering, FFT. etc. Mostly, general purpose rather than application driven architectures are used whenever possible and the available literature is heavily concentrated on the first configuration. At TPD-TNO, the research emphasis is on application driven architectures. and the objectives for the so-called 'matched' architecture designs are: - Capability for a wide range of sizes, starting from small systems. The objective here is design for scalability - Design for systems to be used in harsh environments - Design for minimum connectivity. reduced communication bandwidth, incorporation of dedicated preprocessing. multibus systems, etc. The real-time behaviour of general purpose architectures is not sufficiently predictable and they are not designed to perform acquisition tasks or data-intensive processing with high performance. Matched architectures, on the contrary, are designed for well defined applications and optimized for each application, The key effort in matched architecture research is directed towards efficiently mapping algorithms to processing steps in hardware (and software) architectures. Essentially. the design process is iterative

High-resolution transmission electron microscopy with an electrostatic Zach phase plate

A new method to control lattice-fringe contrast in high-resolution transmission electron microscopy (HRTEM) images by the implementation of a physical phase plate (PP) is proposed. PPs are commonly used in analogy to Zernike PPs in light microscopy to enhance the phase contrast of weak-phase objects with nm-sized features, which often occur in life science applications. Such objects otherwise require strong defocusing, which leads to a degradation of the instrumental resolution and impedes intuitive image interpretation. The successful application of an electrostatic Zach PP in HRTEM is demonstrated by the investigation of single crystalline Si and Ge samples. The influence of the Zach PP on the image formation process is assessed by analyzing the amplitudes of (111) reflections in power spectra which show a cosine-type dependence on the induced phase shift under certain conditions as predicted by theory

Structure of triosephosphate isomerase from Escherichia coli determined at 2.6 A resolution

The structure of triosephosphate isomerase (TIM) from the organism Escherichia coli has been determined at a resolution of 2.6 A. The structure was solved by the molecular replacement method, first at 2.8 A resolution with a crystal grown by the technique of hanging-drop crystallization from a mother liquor containing the transition-state analogue 2-phosphoglycolate (2PG). As a search model in the molecular replacement calculations, the refined structure of TIM from Trypanosoma brucei, which has a sequence identity of 46% compared to the enzyme from E. coli, was used. An E. coli TIM crystal grown in the absence of 2PG, diffracting to 2.6 A resolution, was later obtained by application of the technique of macro-seeding using a seed crystal grown from a mother liquor without 2PG. The final 2.6 A model has a crystallographic R factor of 11.9%, and agrees well with standard stereochemical parameters. The structure of E. coli TIM suggests the importance of residues which favour helix initiation for the formation of the TIM fold. In addition, TIM from E. coli shows peculiarities in its dimer interface, and in the packing of core residues within the beta-barrel

Overexpression of trypanosomal triosephosphate isomerase in Escherichia coli and characterisation of a dimer-interface mutant.

In this paper, the successful expression of trypanosomal triosephosphate isomerase (TIM) from Trypanosoma brucei brucei to high yield in Escherichia coli, using a T7-polymerase-based expression system, is described. Overexpressed trypanosomal TIM is fully active. The measured physicochemical properties of this recombinant TIM and TIM purified from trypanosomes are indistinguishable. Crystals of recombinant TIM have been grown in the presence of 2.4 M ammonium sulphate under the same conditions as for trypanosomally expressed TIM. The recombinant TIM crystal structure has been refined at 0.23 nm resolution; no differences were detected between this structure and the original crystal structure. A TIM mutant was made in which a unique dimer-interface histidine residue (His47) was changed into an asparagine. This variant ([H47N]TIM) could be expressed and purified to homogeneity by a procedure which was somewhat different from the purification of recombinant wild-type TIM. It is shown that the [H47N]TIM dimer is considerably less stable than wild-type trypanosomal TIM. The catalytic activity of [H47N]TIM is concentration dependent. The dilution-dependent inactivation is reversible. His47 is involved in a water-mediated hydrogen bond with Asp385 of the other subunit. The lower stability of the [H47N]TIM dimer implies that this water-mediated hydrogen bond is important for the stability of the TIM dimer

Selective interaction of glycosomal enzymes from Trypanosoma brucei with hydrophobic cyclic hexapeptides.

Hydrophobic cyclic hexapeptides have been reported to selectively inhibit glycosomal triosephosphate isomerase from Trypanosoma brucei (Kuntz et al, 1992, Eur. J. Biochem., 207, 441-447). Here it is shown that this inhibition is not due to a specific interaction between the enzyme and soluble hydrophobic cyclic hexapeptides, but that it is the result of a coprecipitation of trypanosome triosephosphate isomerase with cyclic hexapeptides when the solubilities of the latter are exceeded. A study of the interaction of these hexapeptides with other glycosomal enzymes revealed that several of them, such as phosphoglycerate kinase and hexokinase, also coprecipitated with these peptides, whereas most of the homologous enzymes from other organisms did not coprecipitate, nor were they inactivated

An Interface Point-mutation Variant of Triosephosphate Isomerase Is Compactly Folded and Monomeric At Low-protein Concentrations

Wild-type trypanosomal triosephosphate isomerase (wtTIM) is a very tight dimer, The interface residue His-47 of wtTIM has been mutated into an asparagine, Ultracentrifugation data show that this variant (H47N) only dimerises at protein concentrations above 3 mg/ml, H47N has been characterised at a protein concentration,where it is predominantly a monomer, Circular dichroism measurements in the near-UV and far-UV show that this monomer is a compactly folded protein with secondary structure similar as in wtTIM. The thermal stability of the monomeric H47N is decreased compared to wtTIM: temperature gradient gel electrophoresis (TGGE) measurements give T-m-values of 41 degrees C for wtTIM, whereas the T-m-value for the monomeric form of H47N is approximately 7 degrees C lower