Studies on insect fibrous proteins: the structural protein of the ootheca in the praying mantis, Sphodromatis centralis Rehn

Histological studies have been made on the left colleterial gland which secretes the α-type structural protein in the ootheca of Sphodromantis centralis. Three distinct types of secretory cell are present in the gland wall, and each has a complex apical ‘end-apparatus’ composed of closely packed canalicules radiating from a central cavity. Structural protein is synthesized in the cytoplasm, passed through the canalicules into the cavity of the end-apparatus, and extruded into the lumen as strongly birefringent, apparently membrane-free globules. At this stage the globules consist of numerous concentric layers of long, parallel fibrils, 0·05–0·1 μm in diameter. In each successive layer the fibril direction is rotated through an angle of about 18° about an axis perpendicular to the layers, in the manner of cholesteric phase systems. When sectioned, the globules reveal a regular ‘lamellated’ structure similar to that apparent in sections of some insect and crustacean cuticles. A three-dimensional model has been constructed to illustrate this phenomenon. During ootheca formation fibrils become transformed into thin ‘crystalline’ ribbons, about 15 μm long, 1–2 μm wide and 200–300 Å thick. Very regular diamond-shaped ribbons may be obtained in vitro by mixing structural protein globules from dissected left glands with optimum concentrations (0·005–·025 m ) of calcium chloride in unbuffered solution. Experiments suggest that calcium ions play an important part in the natural process of ribbon formation.     

Automating the identification and analysis of protein {beta}-barrels

ßBarrels are widespread and well-studied featuresof a great many protein structures. In this paper an unsuper-visedmethod for the detection of P-barrels is developed based ontechniques from graph theory. The hydrogen bonded connectivityof ß-sheets is derived using standard pattern recognitiontechniques and expressed as a graph. Barrels correspond to topologicalrings in these connectivity graphs and can thus be identifiedusing ring perception algorithms. Following from this, the characteristictopological structure of a barrel can be expressed using a novelform of reduced nomenclature that counts sequence separationsbetween successive members of the ring set These techniquesare tested by applying them to the detection of barrels in anon-redundant subset of the Brookhaven database. Results indicatethat topological rings do seem to correspond uniquely to ß-barrelsand that the technique, as implemented, finds the majority ofbarrels present in the dataset.     

Stepwise transplantation of an active site loop between heat-labile enterotoxins LT-II and LT-I and characterization of the obtained hybrid toxins

Members of the cholera toxin family, including Escherichia coli heat-labile enterotoxins LT-I and LT-II, catalyze the covalent modification ofintracellular proteins by transfer of ADP-ribose from NAD to a specificarginine of the target protein. The ADP-ribosylating activity of thesetoxins is located in the A-subunit, for which LT-I and LT-II share a 63%sequence identity. The flexible loop in LT-I, ranging from residue 47 to56, closes over the active site cleft. Previous studies have shown thatpoint mutations in this loop have dramatic effects on the activity of LT-I.Yet, in LT-II the sequence of the equivalent loop differs at four positionsfrom LT-I. Therefore five mutants of the active site loop were created by astepwise replacement of the loop sequence in LT-I with virtually all thecorresponding residues in LT- II. Since we discovered that LT-II had noactivity versus the artificial substratediethylamino-benzylidine-aminoguanidine (DEABAG) while LT-I does, ouractive site mutants most likely probe the NAD binding, not the argininebinding region of the active site. The five hybrid toxins obtained (Q49A,F52N, V53T, Q49V/F52N and Q49V/F52N/V53T) show (i) great differences inholotoxin assembly efficiency; (ii) decreased cytotoxicity in Chinesehamster ovary cells; and (iii) increased in vitro enzymatic activitycompared with wild type LT-I. Specifically, the three mutants containingthe F52N substitution display a greater Vmax for NAD than wild type LT-I.The enzymatic activity of the V53T mutant is significantly higher than thatof wild type LT-I. Apparently this subtle variation at position 53 isbeneficial, in contrast to several other substitutions at position 53 whichpreviously had been shown to be deleterious for activity. The most strikingresult of this study is that the active site loop of LT- I, despite greatsensitivity for point mutations, can essentially be replaced by the activesite loop of LT-II, yielding an active 'hybrid enzyme' as well as 'hybridtoxin'.     

Arcing phenomenon as related to fire investigation

A mathematical analysis of arcing phenomenon is presented as related to fire. It is shown that arcing has a great destructive power while a short circuit has not. Experimental results are also given. Note: Dr. Béland, ing., is a Professor of Electrical Engineering at the University of Sherbrooke, Sherbrooke, Quebec, Canada, and conducts research on fire, particularly fires of electrical origin.     

Binding of an arm repeat protein to the kinase domain of the S-locus receptor kinase

Screening of a yeast two-hybrid library for proteins that interact with the kinase domain of an S-locus receptor kinase (SRK) resulted in the isolation of a plant protein called ARC1 (Arm Repeat Containing). This interaction was mediated by the C-terminal region of ARC1 in which five arm repeat units were identified. Using the yeast two-hybrid system and in vitro binding assays, ARC1 was found to interact specifically with the kinase domains from SRK-910 and SRK-A14 but failed to interact with kinase domains from two different Arabidopsis receptor-like kinases. In addition, treatment with a protein phosphatase or the use of a kinase-inactive mutant reduced or abolished the binding of ARC1 to the SRK-910 kinase domain, indicating that the interaction was phosphorylation dependent. Lastly, RNA blot analysis revealed that the expression of ARC1 is restricted to the stigma, the site of the self-incompatibility response.     

Traffic routing in a computer network

This paper presents a technique for tuning Decnet networks so as to minimize the average end-to-end packet delay for the entire network. It represent an adaptation of theoretical work on network flows to the realistic case of Decnet. Using this technique to configure hypothetical networks, it then discusses the behaviour of the Decnet routing algorithm with respect to network size, topological connectivity and traffic configuration.