Crystal structure studies and inhibition kinetics of tripeptide chloromethyl ketone inhibitors with Streptomyces griseus protease B

The bacterial serine protease, SGPB, was inhibited by two specific tripeptide chloromethyl ketones, N-t-butyloxycarbonyl-l-alanylglycyl-l-phenylalanine chloromethyl ketone (BocAGFCK) and N-t-butyloxycarbonyl-glycyl-l-leucyl-l-phenylalanine chloromethyl ketone (BocGLFCK). Crystals of the inhibited complexes were grown and examined by X-ray crystallographic methods. The peptide backbone of each inhibitor is bound by three hydrogen bonds to the main chain of residues Ser214 to Gly216. There are two well-characterized hydrophobic pockets, S₁ and S₂, on the surface of SGPB which accommodate the P₁ and P₂ side-chains of the BocGLFCK inhibitor. A conformational change of Tyr171 is induced by the binding of this inhibitor. Both inhibitors make two covalent bonds to the SGPB enzyme. The imidazole ring of His57 is alkylated at the N^?2 atom and O^γ of Ser195 forms a hemiketal bond with the carbonyl-carbon atom of the inhibitor. Comparison of the binding modes of the two tripeptides in conjunction with the differences in their inhibition constants (K_I) allows one to estimate the binding energy of the leucyl side-chain as ?2.6 kcal mol^?1. The importance of an electrophilic component in the serine protease mechanism, which involves the polarization of the susceptible carbonyl bond of a substrate or inhibitor by the peptide NH groups of Gly193 and Ser195 is discussed.     

Die Rezeptoren an den ersten Antennen vonLeptestheria dahalacensis Rüppel (Crustacea,Conchostraca)

Zusammenfassung Bei dem ConchostracenLeptestheria dahalacensis kommen auf den ersten Antennen etwa 600 gleich aussehende Sinneshaare vor, die in Gruppen von jeweils 25–30 zusammengefaßt sind. Diese Sinneshaare sind in zwei Teile gegliedert, die durch das lichtmikroskopisch gut sichtbare Basalstück (basal bead) voneinander getrennt sind. Dieses bildet die Basis des Haares, dessen Wand im wesentlichen aus Epicuticula besteht. Apikal wird das Haar durch das Endkügelchen (terminal pellet) abgeschlossen. Das Basalstück wird von der untersten Lage der Epicuticula gebildet. Die 4–10 Receptorcilien, die jeweils einzeln ebensovielen Dendriten aufsitzen, ziehen aus dem inneren Teil des Rezeptors, der von insgesamt 5 Hüllzellen umgeben wird, durch das Basalstück, in dem sie stark eingeengt werden und verzweigen sich dann im äußeren Teil des Rezeptors. Sie ziehen bis zum Endkügelchen, in das sie durch einen Porus, den man als Häutungsporus ansprechen kann, eintreten. In der Häutungsvorbereitung wird der Haarbalg von der Hüllzelle 5, das Basalstück von der Hüllzelle 4, der Haarschaft dagegen von der Hüllzelle 3 gebildet. Dabei spaltet sich die Hüllzelle 3 ringspaltförmig auf, so daß in diesem Spalt der neuangelegte Haarschaft handschuhfingerförmig eingestülpt liegt. Die Hüllzelle 2 formt die Spitze des neuen Haares, während die Dendritenscheide von der Hüllzelle 1 abgegeben wird.

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The receptors on the first antennae ofLeptestheria dahalacensis Rüppel (Crustacea, Conchostraca)

Summary On the antennulae ofLeptestheria dahalacensis (Conchostraca) nearly 600 sensory setae of one type are found. They are gathered in groups of 25–30. The single sensory seta is divided into two parts by the basal bead which is easily visible in the light microscope. The basal bead is the socket of the seta, whose wall is mainly built up by the epicuticle. The terminal pellet closes the tip of the seta. The basal bead is derived from the innermost layer of the epicuticle. 4–10 dendrites each with one receptorcilium innervate the receptor. The receptorcilia stretch through the interior part of the receptor and the basal bead into the exterior part, where they branch. They enter the terminal pellet in a porus, which seems to be a moulting porus. The interior part of the receptor is surrounded by 5 sheath cells. During the premoult it becomes obvious, that the socket of the seta is built by the sheath cell 5, the basal bead by the sheath cell 4 and the shaft by the sheath cell 3. For this the sheath cell 3 is divided into two parts. Between this two parts the newly formed cuticle is invaginated. The sheath cell 2 formes the tip and the sheath cell 1 the cuticular sheath of the new bristle.

     

Effect of elevated temperatures on spindle microtubules and chromosome movements in cultured newt lung cells

The effect of elevated temperature shocks (ETS) on anaphase chromosome movements and spindle structure was studied in lung epithelial cells cultured from the newt Taricha granulosa granulosa. Mitosis proceeds normally up to temperatures of 31--32 degrees C. At slightly higher temperatures (33--34 degrees C) anaphase is desynchronized while still higher temperatures (35--36 degrees C) arrest chromosome movements. The desynchronization of chromosome movement is related on the ultrastructural level to the re-arrangement of kinetochore fibre microtubules (Mts) into hexagonally packed structures, and with the disappearance of non-kinetochore Mts. During desynchronized movement the distance a chromosome travels depends on the structure of its kinetochore fibre: those which have divergent kinetochore fibre Mts approach the poles, while those with hexagonally packed kinetochore fibre Mts do not. The data provide evidence concerning the anchorage of Mts and demonstrate that normal anaphase chromosome movements are functionally related both to the rearrangement of kinetochore Mts and to Mt disassembly.     

Selection signatures in Shetland ponies

Shetland ponies were selected for numerous traits including small stature, strength, hardiness and longevity. Despite the different selection criteria, Shetland ponies are well known for their small stature. We performed a selection signature analysis including genome‐wide SNPs of 75 Shetland ponies and 76 large‐sized horses. Based upon this dataset, we identified a selection signature on equine chromosome (ECA) 1 between 103.8 Mb and 108.5 Mb. A total of 33 annotated genes are located within this interval including the IGF1R gene at 104.2 Mb and the ADAMTS17 gene at 105.4 Mb. These two genes are well known to have a major impact on body height in numerous species including humans. Homozygosity mapping in the Shetland ponies identified a region with increased homozygosity between 107.4 Mb and 108.5 Mb. None of the annotated genes in this region have so far been associated with height. Thus, we cannot exclude the possibility that the identified selection signature on ECA1 is associated with some trait other than height, for which Shetland ponies were selected.     

Structural and ecophysiological adaptations to forest gaps

To survive new microclimatic conditions of a forest gap environment, plant species must physiologically and structurally adjust. A morpho-anatomical, ultrastructural and ecophysiological study was performed at three different times in a forest gap that was created by illegal selective logging. The study followed the early successional Actinostemon verticillatus and the late-successional Metrodorea brevifolia, to elucidate the adaptive strategies of acclimation to gaps. Additionally, Schinus terebinthifolius was included in the study in order to test the plasticity of a pioneer species that grows on forest edges, where this species had higher values of leaf thickness, leaf mass area and succulence. M. brevifolia had succulent leaves, high leaf area and a thin cuticle. A. verticillatus presented the densest leaves and was the only species to show leaf morpho-anatomical plasticity. Ultrastructural and physiological differences were observed only in A. verticillatus and M. brevifolia leaves from the gap: increase in the stroma volume, oil droplets, plastoglobuli, photochemical and non-photochemical quenching. Photosynthetic efficiency showed that the early stages of gap formation are the most critical. Acclimation strategies of A. verticillatus suggest this species invests in the efficiency of photosynthesis by increasing its leaf thickness, leaf mass area and in water content maintenance by increasing the density of its leaves, at the expense of gas exchange, was compensated by a high density of stomata. M. brevifolia compensates for the higher cost of leaves and lower leaf plasticity with ultrastructural changes that are used to adjust the photosynthetic process, which promotes a shorter leaf payback time.     

Cytokine gene polymorphisms are associated with risk of urinary bladder cancer and recurrence after BCG immunotherapy

The association of interleukin-1β (IL-1B) -511C?>?T and IL-1 receptor antagonist (IL-1RN) VNTR, transforming growth factor-β (TGF-B1) +28C?>?T and interferon-γ (IFN-G)?+?874T>A polymorphisms with bladder cancer (CaB) susceptibility and risk of recurrence in Bacillus Calmette–Guérin (BCG)-treated patients was analyzed in 287 controls and 213 CaB patients (73 BCG treated). Increased risk was observed with the IL-1RN*2 allele (odds ratio (OR) 5.01) and the IFN-G +874 A allele (OR 1.78). TGF-B TT and IFN-G +874 A carriers were associated with reduced (hazard ratio (HR) 0.37) and enhanced (HR 2.24) risk of recurrence after BCG immunotherapy, respectively. The study suggests that cytokine gene variants may modulate CaB susceptibility and risk of recurrence after BCG immunotherapy.     

Life‐history constraints in grassland plant species: a growth‐defence trade‐off is the norm

Plant growth can be limited by resource acquisition and defence against consumers, leading to contrasting trade‐off possibilities. The competition‐defence hypothesis posits a trade‐off between competitive ability and defence against enemies (e.g. herbivores and pathogens). The growth‐defence hypothesis suggests that strong competitors for nutrients are also defended against enemies, at a cost to growth rate. We tested these hypotheses using observations of 706 plant populations of over 500 species before and following identical fertilisation and fencing treatments at 39 grassland sites worldwide. Strong positive covariance in species responses to both treatments provided support for a growth‐defence trade‐off: populations that increased with the removal of nutrient limitation (poor competitors) also increased following removal of consumers. This result held globally across 4 years within plant life‐history groups and within the majority of individual sites. Thus, a growth‐defence trade‐off appears to be the norm, and mechanisms maintaining grassland biodiversity may operate within this constraint.