Expression of the novel protein PTPIP51 in rat liver: an immunohistochemical study

Expression of the novel protein tyrosine phosphatase interacting protein 51 (PTPIP51) was investigated on mRNA and protein level in the liver of adult Wistar rats. The presence of PTPIP51 mRNA was detected by Northern blotting. Immunostaining showed expression of PTPIP51 protein in distinct non-parenchymal cells. These cells were identified as Kupffer cells, stellate cells and natural killer cells by detection of cell-specific antigens. Whereas most endothelial cells lining large vessels reacted positive to the PTPIP51 antibody, sinusoidal endothelium showed no detectable amount of PTPIP51. Furthermore, PTPIP51 was also found to be expressed in cells forming the biliary tree. An additional subcellular analysis of the non-parenchymal cells by means of electron microscopy showed the presence of PTPIP51 protein in the cytoplasm and in the nuclei of non-parenchymal cells. Most of the hepatocytes did not show any immuno-detectable amount of PTPIP51, yet, some revealed PTPIP51 protein either in the cytoplasm or in the nucleus. This work is dedicated to Professor emeritus D. Sasse, Institute of Anatomy, University of Basel, Switzerland.     

Evaluation of in vitro aldose redutase inhibitory activity of 5-arylidene-2,4-thiazolidinediones

A number of 5-arylidene-2,4-thiazolidinediones containing a hydroxy or a carboxymethoxy group in their 5-benzylidene moiety have been synthesised and evaluated as in vitro aldose reductase (ALR2) inhibitors. Most of them exhibited strong inhibitory activity, with IC(50) values in the range between 0.20 and 0.70 microM. Molecular docking simulations into the ALR2 active site highlighted that the phenolic or carboxylic substituents of the 5-benzylidene moiety can favourably interact, in alternative poses, either with amino acid residues lining the lipophilic pocket of the enzyme, such as Leu300, or with the positively charged recognition region of the ALR2 active site.     

Spatial distribution patterns of Rhynchostegium megapolitanum at the landscape scale ‐ an expanding species?

Question: Is Rhynchostegium megapolitanum an expanding species? Location: Viennese Basin (120‐220 m a.s.1.), Austria. Methods: 121 dry grasslands, were investigated for the occurrence of R. megapolitanum. Nineteen environmental variables at 50 randomly selected sites, species composition at sites with and without R. megapolitanum and the spatial patterns of distribution of the species at the landscape scale were analysed. We compared actual distribution data of three rare species (Didymodon acutus, Pleurochaete squarrosa R. megapolitanum) and a common one (Brachythecium rutabulum) with the distribution obtained by vouchers that were collected between 1860 and 1940 in the investigated area. We calculated a GIS based model pattern and compared it with the actual distribution. Results: R. megapolitanum was detected in 28 of these sites, almost 50 % of its populations produced sporophytes. We found significant differences between sites with and without R. megapolitanum with regard to grassland size, the percentage of silt and of sand in the soil. There were fewer occurrences of historic herbarium vouchers of R. megapolitanum than our current field survey discovered. The GIS based analyses of distribution patterns at the landscape scale showed a clustering of sites in which R. megapolitanum was present or absent. Simulations with a spatially realistic expansion model showed high similarities to the actual distribution of the species. Conclusions: All these analyses suggest that R. megapolitanum has been expanding in the investigated area. A significant increase in temperature and nitrogen deposition within the last hundred years might be the underlying cause for the species' spread.     

Combination of scoring schemes for protein docking

Background  

Docking algorithms are developed to predict in which orientation two proteins are likely to bind under natural conditions. The currently used methods usually consist of a sampling step followed by a scoring step. We developed a weighted geometric correlation based on optimised atom specific weighting factors and combined them with our previously published amino acid specific scoring and with a comprehensive SVM-based scoring function.     

Contributions of the protein environment to the midpoint potentials of the A1 phylloquinones and the Fx iron-sulfur cluster in photosystem I

Electrostatic calculations have predicted that the partial negative charge associated with D575PsaB plays a significant role in modulating the midpoint potentials of the A1A and A1B phylloquinones in photosystem I. To test this prediction, the side chain of residue 575PsaB was changed from negatively charged (D) to neutral (A) and to positively charged (K). D566PsaB, which is located at a considerable distance from either A1A or A1B, and should affect primarily the midpoint potential of FX, was similarly changed. In the 575PsaB variants, the rate of electron transfer from A1A to FX is observed to decrease slightly according to the sequence D/A/K. In the 566PsaB variants, the opposite effect of a slight increase in the rate is observed according to the same sequence D/A/K. These results are consistent with the expectation that changing these residues will shift the midpoint potentials of nearby cofactors to more positive values and that the magnitude of this shift will depend on the distance between the cofactors and the residues being changed. Thus, the midpoint potentials of A1A and A1B should experience a larger shift than will FX in the 575PsaB variants, while FX should experience a larger shift than will either A1A or A1B in the 566PsaB variants. As a result, the driving energy for electron transfer from A1A and A1B to FX will be decreased in the former and increased in the latter. This rationalization of the changes in kinetics is compared with the results of electrostatic calculations. While the altered amino acids shift the midpoint potentials of A1A, A1B, and FX by different amounts, the difference in the shifts between A1A and FX or between A1B and FX is small so that the overall effect on the electron transfer rate between A1A and FX or between A1B and FX is predicted to be small. These conclusions are borne out by experiment.     

Isolation and biochemical characterization of grape malic enzyme

Malic enzyme (ME=L-malate: NADP oxidoreductase; E.C. 1.1.1.40) was extracted by Triton X-100-induced resolubilization of enzyme proteins which denaturize spontaneously upon homogenization of grape berry material. The purification procedure included fractionating with (NH₄)₂SO₄, preparative IEF, and Sephadex G-100 chromatography. ME was identified by TLC of the radioactive product after supplementing the assay mixture with [¹⁴C]malate. Cofactor dependence, pH-optimum and affinities for substrates and cosubstrates were determined. Enzymic pI was found to be 5.8, the Hill coefficients range from 1 to 3. In malate decarboxylating direction at pH 7.4, grape ME displayed positive cooperativity toward the substrate, the curve approaching normal Michaelis-Menten-kinetics at pH 7.0. Substituting Mn²⁺ for Mg²⁺ not only increased maximal turnover rates, but also enzymic affinity for malate. These features were considered indicative of the regulatory properties of the enzyme. Their relevance for grape malate metabolism and fruit ripening is discussed.Abbreviations EDTA ethylenediaminetetraacetic acid - IFF isoelectric focusing - MDH malate dehydrogenase - ME malic enzyme - OAA oxaloacetic acid - PAG polyacrylamide gel - TCA trichloroacetic acid - TLC thin layer chromatography     

An integrated,spatio‐temporal modelling framework for analysing biological invasions

Aim

We develop a novel modelling framework for analysing the spatio‐temporal spread of biological invasions. The framework integrates different invasion drivers and disentangles their roles in determining observed invasion patterns by fitting models to historical distribution data. As a case study application, we analyse the spread of common ragweed (Ambrosia artemisiifolia).

Location

Central Europe.

Methods

A lattice system represents actual landscapes with environmental heterogeneity. Modelling covers the spatio‐temporal invasion sequence in this grid and integrates the effects of environmental conditions on local invasion suitability, the role of invaded cells and spatially implicit “background” introductions as propagule sources, within‐cell invasion level bulk‐up and multiple dispersal means. A modular framework design facilitates flexible numerical representation of the modelled invasion processes and customization of the model complexity. We used the framework to build and contrast increasingly complex models, and fitted them using a Bayesian inference approach with parameters estimated by Markov chain Monte Carlo (MCMC).

Results

All modelled invasion drivers codetermined the A. artemisiifolia invasion pattern. Inferences about individual drivers depended on which processes were modelled concurrently, and hence changed both quantitatively and qualitatively between models. Among others, the roles of environmental variables were assessed substantially differently subject to whether models included explicit source‐recipient cell relationships, spatio‐temporal variability in source cell strength and human‐mediated dispersal means. The largest fit improvements were found by integrating filtering effects of the environment and spatio‐temporal availability of propagule sources.

Main conclusions

Our modelling framework provides a straightforward means to build integrated invasion models and address hypotheses about the roles and mutual relationships of different putative invasion drivers. Its statistical nature and generic design make it suitable for studying many observed invasions. For efficient invasion modelling, it is important to represent changes in spatio‐temporal propagule supply by explicitly tracking the species’ colonization sequence and establishment of new populations.

     

Identification and characterization of an IgG binding protein in the secretion of the rat coagulating gland

A merocrine released protein (named 115k protein) was highly enriched from the secretion of the rat coagulating gland. The protein has a molecular mass of 115 kDa as calculated by SDS-PAGE under reducing conditions. Furthermore, the 115 kDa protein is glycosylated, and carries Man, GlcNAc, Gal, Fuc and sialic acid residues. For identification, N-terminal amino acid and nucleotide sequence analyses were performed. The sequences obtained showed 86 to 100% identity with human and mouse IgGFc binding proteins. The functional capacity of IgG binding of the 115 kDa protein was shown by overlay experiments, indicating its membership in the IgG binding protein family.