Reversible cross-linking of crystalline bacterial surface layer glycoproteins through their glycan chains

After periodate oxidation and incubation with dithiodipropionic acid dihydrazide cross-linking of the crystalline surface layer (S-layer) glycoproteins of Clostridium thermohydrosulfuricum L111-69 and Bacillus alvei CCM 2051 was achieved specifically through the glycan chains. The cross-linked S-layers were used for the immobilization of chemically synthesized, spacer-linked, tumour-associated T-disaccharide [Gal(13)GalNAc]. Electron microscopical evaluation of the resulting conjugates showed densely packed, multilayered S-layer structures loaded with the immobilized ligand. After reductive cleavage of the disulphide bond of dithiodipropionic acid by dithiothreitol, monomeric haptenated S-layer conjugates could be obtained. Both the cross-linked and the monomeric type of conjugate might be useful for assessment of specific immune responses, which, in general, can be elicited by those artificial antigens. Correspondence to: P. Messner     

Immuno- and constitutive proteasome crystal structures reveal differences in substrate and inhibitor specificity

Constitutive proteasomes and immunoproteasomes shape the peptide repertoire presented by major histocompatibility complex class I (MHC-I) molecules by harboring different sets of catalytically active subunits. Here, we present the crystal structures of constitutive proteasomes and immunoproteasomes from mouse in the presence and absence of the epoxyketone inhibitor PR-957 (ONX 0914) at 2.9 ? resolution. Based on our X-ray data, we propose a unique catalytic feature for the immunoproteasome subunit β5i/LMP7. Comparison of ligand-free and ligand-bound proteasomes reveals conformational changes in the S1 pocket of β5c/X but not β5i, thereby explaining the selectivity of PR-957 for β5i. Time-resolved structures of yeast proteasome:PR-957 complexes indicate that ligand docking to the active site occurs only via the reactive head group and the P1 side chain. Together, our results support structure-guided design of inhibitory lead structures selective for immunoproteasomes that are linked to cytokine production and diseases like cancer and autoimmune disorders.     

Experimental ‘omics’ data in tree research: facing complexity

High-throughput experimental technology has provided insight into the inner functioning of plants. The current experimental technology facilitates the study of plant systems in a holistic manner, measuring observables from the genome, proteome, and metabolome up to the level of the ecosystem. The call for a systemic view in plant research is being made from multiple research fields. Although not yet fully developed for tree research, data sources are also rapidly growing in this area. Nevertheless, there are challenges and pitfalls in dealing with such increases in data. Some of these difficulties are deeply rooted in the complexity of the evolutionary systems. The lessons from complexity theory are rooted in studies performed several decades ago. Honouring principles that were formulated before bioinformatics and systems biology had been introduced facilitates the derivation of analytical methods with the potential to overcome these challenges in several ways.     

Diurnal periodicity of chalcone-synthase activity during the development of oat primary leaves

Chalcone-synthase (CHS) activity was followed during the development of primary leaves of oat (Avena sativa L.) seedlings grown under different illumination conditions. Continuous darkness and continuous light resulted in similar time courses of enzyme activity. The maximum of CHS activity in etiolated leaves was delayed by 1 d and reached about half the level of that of light-grown leaves. In seedlings grown under defined light-dark cycles a diurnal rhythm of CHS activity and its protein level was observed which followed the rhythm of CHS-mRNA translational activity (Knogge et al. 1986). This rhythm persisted in continuous light after a short-term pre-exposure to the light-dark cycle but not in continuous darkness.Abbreviations CHS chalcone synthase - PAL phenylalanine ammonio lyase Financial support by the Deutsche Forschungsgemeinschaft is gratefully acknowledged (G.W., We 630/9-7; We 630/10-1). Thanks are given to Dr. St. Kellam (Department of Plant Microbiological Sciences, University of Canterbury, New Zealand) for correcting the English.     

Circadian locomotor activity of Musca flies: Recording method and effects of 10 Hz square-wave electric fields

Musca domestica flies that were exposed to a uniform vertical 10 Hz electric square-wave field of 1 kVm⁻¹ changed the period length of their circadian locomotor activity rhythm. Under constant conditions, the clock of short-period flies was slowed down by the field, whereas the clock of long-period flies either was affected only scarcely (experiments at about 19°C) or ran faster (experiments at 25°C). If the field was applied for only 12 h daily, then 30–40% of the flies were synchronized. Thus, the field could function as a weak “Zeitgeber” (synchronizer). If the field was increased to 10 kVm⁻¹, then 50–70% of the flies were synchronized. Flies avoided becoming active around the onset of the 12 h period of exposure to a 10 Hz field. The results of these experiments are discussed with respect to similar experiments by Wever on the effects of exposure to a 10 Hz field on the circadian system of man. © 1996 Wiley-Liss, Inc.     

Zur Beurteilung der Feldresistenz von Wintergerste (Hordeum vulgare L.) gegenüber Zwergrost (Puccinia hordei Otth)

Eine anorganische Phosphatase aus Escherichia coli wurde als neuer Enzym‐Marker zum ELISA‐Test bei vier Nepoviren‐raspberry ringspot virus, strawberry latent ringspot virus, tomato black ring virus und arabis mosaic virus‐benutzt. Ein Vergleich des ELISA bei Nutzung von Peroxydase aus Meerrettich und von anorganischer Pyrophosphatase als Marker ergab, daß die Nutzung von Pyrophosphatase einen empfindlicheren Nachweis der Viren in gereinigten Präparaten, in Extrakten aus krautigen Testpflanzen sowie in Rohsäften von Himbeerpflanzen erlaubte.     

Accuracy of prediction equations for determining one repetition maximum bench press in women before and after resistance training

Repetitions to fatigue (RTF) using less than a 1 repetition maximum (1RM) load (RepWt) have been shown to be a good predictor of 1RM strength in men, but such information is scarce in women. The purpose of this study was to evaluate the accuracy of current prediction equations to estimate 1RM bench press performance and to determine whether resistance training changes the capability to predict 1RM from muscular endurance repetitions in young women. Members (n = 103) of a required wellness course were measured for 1RM bench press and RTF using randomly assigned percentages between 60% and 90% of the 1RM (RepWt) before and after 12 weeks of progressive resistance training. The %1RM used to perform RTF remained the same for each individual after training (75.6% +/- 10.3%) as before. One repetition maximum bench press increased significantly after training (28% +/- 21%). Although the change in the group average for RTF (0.6 +/- 6.1) was not significant, the correlation between pretraining and posttraining RTF was moderate (r = 0.66; p < 0.01), and individual differences in percentage change in RTF were substantial (27% +/- 99%). The percentage change in 1RM was not significantly related to initial 1RM (r = -0.05), but it was negatively related to the change in RTF (r = -0.40; p < 0.01). Prediction equations were more accurate in the pretraining and posttraining conditions, in which fewer than 10 RTF were used. Resistance training may alter the relationship between strength and muscle endurance across a wide range of RTF in young women without compromising the accuracy of predicting maximal strength.