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1.
Liang Hong Melissa?A. Sharp Simón Poblete Ralf Biehl Michaela Zamponi Noemi Szekely Marie-Sousai Appavou Roland?G. Winkler Rachel E. Nauss Alexander Johs Jerry?M. Parks Zheng Yi Xiaolin Cheng Liyuan Liang Michael Ohl Susan?M. Miller Dieter Richter Gerhard Gompper Jeremy?C. Smith 《Biophysical journal》2014,107(2):393-400
The functional efficacy of colocalized, linked protein domains is dependent on linker flexibility and system compaction. However, the detailed characterization of these properties in aqueous solution presents an enduring challenge. Here, we employ a novel, to our knowledge, combination of complementary techniques, including small-angle neutron scattering, neutron spin-echo spectroscopy, and all-atom molecular dynamics and coarse-grained simulation, to identify and characterize in detail the structure and dynamics of a compact form of mercuric ion reductase (MerA), an enzyme central to bacterial mercury resistance. MerA possesses metallochaperone-like N-terminal domains (NmerA) tethered to its catalytic core domain by linkers. The NmerA domains are found to interact principally through electrostatic interactions with the core, leashed by the linkers so as to subdiffuse on the surface over an area close to the core C-terminal Hg(II)-binding cysteines. How this compact, dynamical arrangement may facilitate delivery of Hg(II) from NmerA to the core domain is discussed. 相似文献
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Richard Ward Christos Pliotas Emma Branigan Christian Hacker Akiko Rasmussen Gregor Hagelueken Ian R. Booth Samantha Miller John Lucocq James H. Naismith Olav Schiemann 《Biophysical journal》2014
Mechanosensitive channel proteins are important safety valves against osmotic shock in bacteria, and are involved in sensing touch and sound waves in higher organisms. The mechanosensitive channel of small conductance (MscS) has been extensively studied. Pulsed electron-electron double resonance (PELDOR or DEER) of detergent-solubilized protein confirms that as seen in the crystal structure, the outer ring of transmembrane helices do not pack against the pore-forming helices, creating an apparent void. The relevance of this void to the functional form of MscS in the bilayer is the subject of debate. Here, we report PELDOR measurements of MscS reconstituted into two lipid bilayer systems: nanodiscs and bicelles. The distance measurements from multiple mutants derived from the PELDOR data are consistent with the detergent-solution arrangement of the protein. We conclude, therefore, that the relative positioning of the transmembrane helices is preserved in mimics of the cell bilayer, and that the apparent voids are not an artifact of detergent solution but a property of the protein that will have to be accounted for in any molecular mechanism of gating. 相似文献
3.
Emily H. Belser Bradley S. Cohen Shamus P. Keeler Charles H. Killmaster John W. Bowers Karl V. Miller 《PloS one》2015,10(3)
Cranial/intracranial abscess disease is an emerging source of significant mortality for male white-tailed deer (Odocoileus virginianus). Most cases of cranial/intracranial abscess disease are associated with infection by the opportunistic pathogen Trueperella pyogenes although the relationship between the prevalence of the bacteria and occurrence of disease is speculative. We examined 5,612 hunter-harvested deer from 29 sites across all physiographic provinces in Georgia for evidence of cranial abscess disease and sampled the forehead, lingual, and nasal surfaces from 692 deer. We used polymerase chain reaction (PCR) to determine presence of T. pyogenes from these samples. We found T. pyogenes prevalence at a site was a predictor for the occurrence of cranial abscess disease. Prevalence of T. pyogenes did not differ between samples from the nose or tongue although prevalence along the forehead was greater for males than females (p = 0.04), particularly at sites with high occurrence of this disease. Socio-sexual behaviors, bacterial prevalence, or physiological characteristics may predispose male deer to intracranial/cranial abscess disease. Determination of factors that affect T. pyogenes prevalence among sites may help explain the occurrence of this disease among populations. 相似文献
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Human foamy virus (HFV) is a retrovirus of the spumavirus family. We have constructed vectors based on HFV that encode neomycin phosphotransferase and alkaline phosphatase. These vectors are able to transduce a wide variety of vertebrate cells by integration of the vector genome. Unlike vectors based on murine leukemia virus, HFV vectors are not inactivated by human serum, and they transduce stationary-phase cultures more efficiently than murine leukemia virus vectors. These properties, as well as their large packaging capacity, make HFV vectors promising gene transfer vehicles. 相似文献
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Alexander F. Wall Yurena Yanes Joshua H. Miller Arnold I. Miller 《Biodiversity and Conservation》2018,27(2):395-415
Natural areas near human-modified landscapes experience factors that may affect local biodiversity at levels commensurate with natural environmental factors. The land snails of the Canary Islands provide excellent opportunities to evaluate the importance of anthropogenic agents in mediating the diversity and distribution of species. Land snails are particularly sensitive to disturbance and are an integral part of terrestrial ecosystems. This study analyzed the distributions and abundances of terrestrial macrosnail shell assemblages at 60 localities throughout the coastal scrub biome of the Canary Islands. This was accomplished using data on natural and anthropogenic variables to assess their relative importance in governing land snail diversity. A total of 34,801 dead shells represented a diverse malacofauna with highly localized endemism. Due to uncertain species identifications, samples from the 18 sites from the two easternmost islands are described, but excluded from statistical analyses. Regression tree analysis indicated that proximity to agricultural sites was the most important predictor of species diversity. Sites with no or very little agricultural area (≤ 0.167 km2) within a 1 km radius had significantly higher richness and diversity. These results have implications for Canary Islands conservation. Protected areas that are patchworks of natural and agricultural landscapes are still subject to native biodiversity loss because of anthropogenic impacts even when the footprint of agriculture is small. 相似文献
8.
Christine Jean Xiao Lei Chen Ju-Ock Nam Isabelle Tancioni Sean Uryu Christine Lawson Kristy K. Ward Colin T. Walsh Nichol L.G. Miller Majid Ghassemian Patric Turowski Elisabetta Dejana Sara Weis David A. Cheresh David D. Schlaepfer 《The Journal of cell biology》2014,204(2):247-263
Pharmacological focal adhesion kinase (FAK) inhibition prevents tumor growth and metastasis, via actions on both tumor and stromal cells. In this paper, we show that vascular endothelial cadherin (VEC) tyrosine (Y) 658 is a target of FAK in tumor-associated endothelial cells (ECs). Conditional kinase-dead FAK knockin within ECs inhibited recombinant vascular endothelial growth factor (VEGF-A) and tumor-induced VEC-Y658 phosphorylation in vivo. Adherence of VEGF-expressing tumor cells to ECs triggered FAK-dependent VEC-Y658 phosphorylation. Both FAK inhibition and VEC-Y658F mutation within ECs prevented VEGF-initiated paracellular permeability and tumor cell transmigration across EC barriers. In mice, EC FAK inhibition prevented VEGF-dependent tumor cell extravasation and melanoma dermal to lung metastasis without affecting primary tumor growth. As pharmacological c-Src or FAK inhibition prevents VEGF-stimulated c-Src and FAK translocation to EC adherens junctions, but FAK inhibition does not alter c-Src activation, our experiments identify EC FAK as a key intermediate between c-Src and the regulation of EC barrier function controlling tumor metastasis. 相似文献
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