0000000000124638

AUTHOR

Frank Müller

New national and regional bryophyte records, 61

New record of phytogeographical interest, at national or regional scale, for 47 bryophytes taxa are reported. New sites are located in 23 different geographical areas of . In particular, for each taxon, data on taxonomy, ecological as well as phytogeographical features are specified

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Domains of the E1 Protein of Human Papillomavirus Type 33 Involved in Binding to the E2 Protein

Papillomavirus E1 and E2 proteins are essential for the initiation of viral DNA replication. We have now analyzed the interaction of E1 and E2 of human papillomavirus type 33, which is associated with cervical carcinoma. When synthesized in insect cells using the baculovirus expression system, the E1 and E2 proteins interacted efficiently at 4 degree. A monoclonal antibody recognizing E1 amino acids 584--600 inhibited the binding of E2 and vice versa, indicating that these amino acids are involved in E2 binding. To confirm this result, a mutational analysis of E1 was performed. The E2 binding activity of E1 deletion and point mutant proteins was assayed using glutathione S-transferase E1 fu…

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Visualization of a covalent intermediate between microsomal epoxide hydrolase, but not cholesterol epoxide hydrolase, and their substrates

Mammalian soluble and microsomal epoxide hydrolases have been proposed to belong to the family of alpha/beta-hydrolase-fold enzymes. These enzymes hydrolyse their substrates by a catalytic triad, with the first step of the enzymatic reaction being the formation of a covalent enzyme-substrate ester. In the present paper, we describe the direct visualization of the ester formation between rat microsomal epoxide hydrolase and its substrate. Microsomal epoxide hydrolase was precipitated with acetone after brief incubation with [1-(14)C]epoxystearic acid. After denaturing SDS gel electrophoresis the protein-bound radioactivity was detected by fluorography. Pure epoxide hydrolase and crude micros…

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Functional Characterization of a Guanylyl Cyclase-activating Protein from Vertebrate Rods

The membrane-bound guanylyl cyclase in vertebrate photoreceptor cells is one of the key enzymes in visual transduction. It is highly sensitive to the free calcium concentration ([Ca2+]). The activation process is cooperative and mediated by a novel calcium-binding protein named GCAP (guanylyl cyclase-activating protein). We isolated GCAP from bovine rod outer segments, determined amino acid sequences of proteolytically obtained peptides, and cloned its gene. The Ca2+-bound form of native GCAP has an apparent molecular mass of 20.5 kDa and the Ca2+-free form of 25 kDa as determined by SDS-polyacrylamide gel electrophoresis. Recombinant GCAP was functionally expressed in Escherichia coli. Act…

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Quo Vadis, Orthotrichum pulchellum? A Journey of Epiphytic Moss across the European Continent

Orthotrichum pulchellum is a species of epiphytic moss in which a significant expansion from the oceanic part of Europe to the east of the continent has been observed in the recent two decades. The improvement in air quality in Central and Eastern Europe, but also climate change, probably plays a role in this. This study shows what direction of its spreading we can expect in the future. Ecological niche modeling (ENM) is a widespread method to find out species niches in environmental and geographical space, which allows us to highlight areas that have a higher probability of occurrences of the studied species, based on identifying similar environmental conditions to those already known. We …

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Interaction of glutamic-acid-rich proteins with the cGMP signalling pathway in rod photoreceptors.

The assembly of signalling molecules into macromolecular complexes (transducisomes) provides specificity, sensitivity and speed in intracellular signalling pathways. Rod photoreceptors in the eye contain an unusual set of glutamic-acid-rich proteins (GARPs) of unknown function. GARPs exist as two soluble forms, GARP1 and GARP2, and as a large cytoplasmic domain (GARP' part) of the beta-subunit of the cyclic GMP-gated channel. Here we identify GARPs as multivalent proteins that interact with the key players of cGMP signalling, phosphodiesterase and guanylate cyclase, and with a retina-specific ATP-binding cassette transporter (ABCR), through four, short, repetitive sequences. In electron mic…

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New national and regional bryophyte records, 40

1. Aneura pseudopinguis (Herzog) PocsContributor: K. HylanderEthiopia: Kaffa, Bonga, Gimbo, Meligawa, Barta forest, 3 km ENE of Bonga, moist Afromontane forest, among other bryophytes on dead wood,...

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Catalytic triad of microsomal epoxide hydrolase: replacement of Glu404 with Asp leads to a strongly increased turnover rate

Microsomal epoxide hydrolase (mEH) belongs to the superfamily of α/β-hydrolase fold enzymes. A catalytic triad in the active centre of the enzyme hydrolyses the substrate molecules in a two-step reaction via the intermediate formation of an enzyme-substrate ester. Here we show that the mEH catalytic triad is composed of Asp226, Glu404 and His431. Replacing either of these residues with non-functional amino acids results in a complete loss of activity of the enzyme recombinantly expressed in Saccharomyces cerevisiae. For Glu404 and His431 mutants, their structural integrity was demonstrated by their retained ability to form the substrate ester intermediate, indicating that the lack of enzymi…

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