Structural and functional characterization of a novel T cell receptor co-regulatory protein complex, CD97-CD55

CD97, the archetypal member of the EGF-TM7 protein family, is constitutively expressed on granulocytes and monocytes and rapidly up-regulated on T and B cells following activation. The key isoform of CD97 expressed on leukocytes binds the complement regulatory protein CD55 (also termed decay-accelerating factor). CD97 has been shown recently to mediate co-stimulation of T cells via CD55. Here, we demonstrate that blocking the interaction between CD55 on monocytes and CD97 on T cells leads to inhibition of proliferation and interferon-gamma secretion. This implies that bidirectional interactions between CD97 and CD55 are involved in T cell regulation. Structural studies presented here reveal the molecular basis for this activity. We have solved the structure of EMR2, a very close homolog of CD97, using x-ray crystallography. NMR-based chemical shift mapping of the EMR2-CD55 interaction has allowed us to generate a model for the CD97-CD55 complex. The structure of the complex reveals that the T cell and complement regulatory activities of CD55 occur on opposite faces of the molecule. This suggests that CD55 might simultaneously regulate both the innate and adaptive immune responses, and we have shown that CD55 can still regulate complement when bound to CD97.     

Ca2+-dependent interface formation in fibrillin-1

The calcium-binding epidermal growth factor-like (cbEGF) domain is a common structural motif in extracellular and transmembrane proteins. K(d) values for Ca2+ vary from the millimolar to nanomolar range; however the molecular basis for this variation is poorly understood. We have measured K(d) values for six fibrillin-1 cbEGF domains, each preceded by a transforming growth factor beta-binding protein-like (TB) domain. Using NMR and titration with chromophoric chelators, we found that K(d) values varied by five orders of magnitude. Interdomain hydrophobic contacts between TB-cbEGF domains were studied by site-directed mutagenesis and could be correlated directly with Ca2+ affinity. Furthermore, in TB-cbEGF pairs that displayed high-affinity binding, NMR studies showed that TB-cbEGF interface formation was strongly Ca2+-dependent. We suggest that Ca2+ affinity is a measure of interface formation in both homologous and heterologous cbEGF domain pairs, thus providing a measure of flexibility in proteins with multiple cbEGF domains. These data highlight the versatile role of the cbEGF domain in fine tuning the regional flexibility of proteins and provide new constraints for the organization of fibrillin-1 within 10-12-nm microfibrils of the extracellular matrix.     

Detecting spatial patterns in species composition with multiple plot similarity coefficients and singularity measures

Recently, several multiple plot similarity indices have been presented that cure some of the problems associated with the approaches for the calculation of compositional similarity for groups of plots by averaging pairwise similarities. These new indices calculate the similarity between more than two plots whilst considering the species composition on all compared plots. The resulting similarity value is true for the whole group of plots considered (called neighborhood in the following). Here, we review the possibilities for multiple plot similarity calculation and additionally explore coefficients that examine multiple plot similarity between a reference plot (named focal plot in the following) and any number of surrounding plots. The latter represent measures of singularity. Further, we establish a framework for applying these two kinds of multiple plot measures to gridded data including an algorithm for testing the significance of calculated values against random expectations. The capability of multiple plot measures for detecting species compositional gradients and local/regional hotspots within this framework is tested. For this purpose, several artificial data sets with known gradients in species composition (random, gradient, central hotspot, hotspot bottom right) are constructed on the basis of a real data set from a Tundra ecosystem in northern Sweden (Abisko). The coefficients that best reflect the positions of the plots on the realized gradients in species composition are considered as performing best with regard to pattern detection. The tested measures of multiple plot similarity and singularity produced considerably different results when applied to one real and 4 artificial data sets. The newly proposed symmetric singularity coefficient has the best overall performance which makes it suitable for local/regional hotspot detection and for incorporating local to regional similarity analyses in reserve selection procedures.     

Inventory, differentiation, and proportional diversity: a consistent terminology for quantifying species diversity

Almost half a century after Whittaker (Ecol Monogr 30:279–338, 1960) proposed his influential diversity concept, it is time for a critical reappraisal. Although the terms alpha, beta and gamma diversity introduced by Whittaker have become general textbook knowledge, the concept suffers from several drawbacks. First, alpha and gamma diversity share the same characteristics and are differentiated only by the scale at which they are applied. However, as scale is relative––depending on the organism(s) or ecosystems investigated––this is not a meaningful ecological criterion. Alpha and gamma diversity can instead be grouped together under the term “inventory diversity.” Out of the three levels proposed by Whittaker, beta diversity is the one which receives the most contradictory comments regarding its usefulness (“key concept” vs. “abstruse concept”). Obviously beta diversity means different things to different people. Apart from the large variety of methods used to investigate it, the main reason for this may be different underlying data characteristics. A literature review reveals that the multitude of measures used to assess beta diversity can be sorted into two conceptually different groups. The first group directly takes species distinction into account and compares the similarity of sites (similarity indices, slope of the distance decay relationship, length of the ordination axis, and sum of squares of a species matrix). The second group relates species richness (or other summary diversity measures) of two (or more) different scales to each other (additive and multiplicative partitioning). Due to that important distinction, we suggest that beta diversity should be split into two levels, “differentiation diversity” (first group) and “proportional diversity” (second group). Thus, we propose to use the terms “inventory diversity” for within-sample diversity, “differentiation diversity” for compositional similarity between samples, and “proportional diversity” for the comparison of inventory diversity across spatial and temporal scales. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Habitat engineering under dry conditions: The impact of pikas (Ochotona pallasi) on vegetation and site conditions in southern Mongolian steppes

Question: Does ecosystem engineering by small mammals have a significant influence on vegetation patterns in the arid steppe vegetation of southern Mongolia? Location: Gobi Altay Mountains, southern Mongolia. Methods: We assessed the impact of the small lagomorph Ochotona pallasi on plant community composition, nutrient levels and biomass production in montane desert steppes. Data were derived from vegetation relevés, harvests of above‐ground standing crop and a bioassay, followed by analyses of soil and plant nutrient contents. Results: Although the local climate is arid with <150 mm annual precipitation, clear evidence of allogenic ecosystem engineering was found. Plant communities on burrows differed from those on undisturbed steppe in that they contained more species of annuals and dwarf shrubs, and a greater abundance of the important fodder grass Agropyron cristatum. Standing crop and nutrient concentrations were higher for plants growing on burrow soil. In situ measurements and a pot experiment showed that this effect was related to increased levels of soil nutrients (P, K, N) rather than moisture availability. Conclusions: The study confirms that O. pallasi positively influences soil nutrient levels on its burrows, which leads to increased grassland productivity even under dry conditions. Thus, O. pallasi does not deteriorate site conditions, and the need for presently applied pest control schemes aimed at this species should be reassessed.     

Glucose-containing polymer rings enable fed-batch operation in microtiter plates with parallel online measurement of scattered light,fluorescence, dissolved oxygen tension,and pH

Bioprocesses operated in batch mode can induce adverse effects like overflow metabolism, substrate inhibition, osmotic inhibition, oxygen limitation, and catabolite repression. To avoid these adverse effects, fed-batch is the predominant operation mode in industrial production. Nevertheless, screening for optimal production strains is usually performed in microtiter plates and shake flasks operated in batch mode without any online monitoring. Recently, a polymer-based controlled-release fed-batch microtiter plate with stable glucose release characteristics was described. In this study, a glucose-containing polymer matrix was used to manufacture polymer rings that were placed at the bottom of a 48-well microtiter plate. Thereby, the liquid content of the well became accessible for optical measurement by the BioLector device. Reflections caused by the polymer ring were minimized by adjusting the scattered-light measurement position. Influences on the measurement of the dissolved oxygen tension and pH could be avoided by choosing appropriate polymer-ring geometries. These adjustments enabled parallel online measurement of scattered light, fluorescence, dissolved oxygen tension, and pH of Escherichia coli BL21 (DE3) fed-batch cultivations. The online monitoring and fed-batch operation capabilities of the fed-batch microtiter plate presented in this study finds optimal application in screenings and initial process development.