Papers by christoph kempf
Medecine Et Hygiene, 1994
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Kasermann Fabian Jurs Mathias Kempf Christoph Prionen Erkrankungen Beim Menschen Und Die Sicherheit Von Blut Und Plasmatischen Produkten in Seifried Erhard Quo Vadis Plasmaproteine Munchen Gesellschaft Fur Thrombose Und Hamostaseforschung, 2006
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Methods in Enzymology, 1984
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Dynamics of Membrane Assembly, 1992
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Future Virology, 2006
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Clinical Reviews in Allergy and Immunology, 2005
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Journal of Biological Chemistry, May 3, 2001
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Virology, May 31, 1985
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Journal of Virology, Nov 1, 2010
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Viruses, 2016
Parvovirus B19 (B19V) is known as the human pathogen causing the mild childhood disease erythema ... more Parvovirus B19 (B19V) is known as the human pathogen causing the mild childhood disease erythema infectiosum. B19V shows an extraordinary narrow tissue tropism for erythroid progenitor cells in the bone marrow, which is determined by a highly restricted uptake. We have previously shown that the specific internalization is mediated by the interaction of the viral protein 1 unique region (VP1u) with a yet unknown cellular receptor. To locate the receptor-binding domain (RBD) within the VP1u, we analyzed the effect of truncations and mutations on the internalization capacity of the recombinant protein into UT7/Epo cells. Here we report that the N-terminal amino acids 5-80 of the VP1u are necessary and sufficient for cellular binding and internalization; thus, this N-terminal region represents the RBD required for B19V uptake. Using site-directed mutagenesis, we further identified a cluster of important amino acids playing a critical role in VP1u internalization. In silico predictions a...
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Journal of virology, 2016
Although the mechanism is not well understood, growing evidence indicates that the nonenveloped p... more Although the mechanism is not well understood, growing evidence indicates that the nonenveloped parvovirus minute virus of mice (MVM) may actively egress before passive release through cell lysis. We have dissected the late maturation steps of the intranuclear progeny with the aims of confirming the existence of active prelytic egress and identifying critical capsid rearrangements required to initiate the process. By performing anion-exchange chromatography (AEX), we separated intranuclear progeny particles by their net surface charges. Apart from empty capsids (EC), two distinct populations of full capsids (FC) arose in the nuclei of infected cells. The earliest population of FC to appear was infectious but, like EC, could not be actively exported from the nucleus. Further maturation of this early population, involving the phosphorylation of surface residues, gave rise to a second, late population with nuclear export potential. While capsid surface phosphorylation was strictly asso...
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Antivir Res, 1997
Photodynamic reactions induced by singlet oxygen-generating agents are known to inactivate envelo... more Photodynamic reactions induced by singlet oxygen-generating agents are known to inactivate enveloped viruses. In this report we demonstrate that the water-insoluble photosensitizer buckminsterfullerene (C60) can be used to mediate the inactivation of enveloped viruses. Viruses from two different families, Semliki Forest virus (SFV, Togaviridae) and vesicular stomatitis virus (VSV, Rhabdoviridae) were used as model systems. Buffered solutions containing C60 plus either of these viruses were illuminated with visible light for up to 5 h, resulting in a loss of infectivity of more than 7 log10/ml (TCID50). Furthermore, it was demonstrated that this viral inactivation was oxygen-dependent and equally efficient in solutions containing protein. C60 yields singlet oxygen in very high amounts and is completely inert to photo-oxidative destruction. In addition, it can be easily removed and recycled from aqueous solutions. For these reasons, it may prove useful in the inactivation of viruses in biological systems.
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Bioscience Rep, 1988
The mechanism of the processes leading to membrane fusion is as yet unknown. In this report we de... more The mechanism of the processes leading to membrane fusion is as yet unknown. In this report we demonstrate that changes in membrane potential and potassium fluxes correlate with Semliki Forest virus induced cell-cell fusion at mildly acidic pH. The changes observed occur only at pH's below 6.2 corresponding to values required to trigger the fusion process. A possible role of these alterations of the plasma membrane related to membrane fusion phenomena is discussed.
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Bioscience Rep, 1993
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Journal of Biological Chemistry
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Bioconjugate Chemistry, 2015
Viruses are evolutionarily developed cell-entering nanomachines, which are frequently used as gen... more Viruses are evolutionarily developed cell-entering nanomachines, which are frequently used as gene or drug delivery systems. Parvovirus B19 (B19V) shows a remarkably restricted tropism for erythropoietin-dependent erythroid differentiation stages, and thus this virus provides an opportunity to deliver cargo to these intermediate differentiated cells. Here we report the construction of a delivery system from B19V subunits that maintains the highly selective cell-entry of the native virus and offers versatile cargo transport. To obtain this specific carrier, we conjugated the cell-targeting VP1u region of B19V to NeutrAvidin as a loading platform for biotinylated cargos. The VP1u-NeutrAvidin conjugate delivered fluorophores, DNA, and toxic payloads specifically to erythroid cells around the proerythroblast differentiation stage, including erythroleukemic cells. The VP1u-NeutrAvidin represents a unique cell surface marker which exclusively detects intermediate erythroid differentiation stages. Furthermore, the cell-entering property of this viral-based targeting system offers opportunities for erythroid-specific drug delivery or gene therapy.
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Biological Membranes: Structure, Biogenesis and Dynamics, 1994
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Virology, 2004
Minute virus of mice (MVM) infection is disrupted by proteasome inhibitors. Here, we show that in... more Minute virus of mice (MVM) infection is disrupted by proteasome inhibitors. Here, we show that inhibition of the ubiquitin-proteasome pathway did not affect viral entry and had influence neither on the natural proteolytic cleavage of VP2 to VP3 nor on the externalization of the N terminal of VP1. In both MG132-treated and untreated cells, MVM particles accumulated progressively in the perinuclear region. However, in MG132-treated cells, MVM was not able to penetrate into the nuclei, remaining blocked in the perinuclear region without capsid disassembly. MVM was similarly sensitive to MG132 in the two cell lines tested, A9 and NB324K. After releasing from the reversible MG132 block, MVM recovered the ability to translocate to the nuclei and replicate. Analysis of viral capsid proteins during internalization showed no evidence of capsid ubiquitination or degradation. We examined the effect of MG132 on two other parvoviruses, canine (CPV) and bovine parvovirus (BPV). Similarly to MVM, ...
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The Journal of physiology, 1992
1. Cell pairs of an insect cell line (Aedes albopictus, clone C6/36) were used to study the elect... more 1. Cell pairs of an insect cell line (Aedes albopictus, clone C6/36) were used to study the electrical properties of intercellular junctions. A double voltage-clamp approach was adopted to control the voltage gradient between the cells and measure the intracellular current flow. 2. Determinations of junctional conductance (gj) revealed two types of intercellular contacts, gap junctions and cytoplasmic bridges. Identification occurred by means of functional criteria, i.e. the dependency of gj on (i) junctional membrane potential, (ii) non-junctional membrane potential, and (iii) heptanol. 3. In cell pairs with putative gap junctions, gj was dependent on the junctional membrane potential (Vj). When determined at the beginning of voltage pulses, gj was insensitive to Vj; when determined at the end of 15 s pulses, it depended on Vj in a bell-shaped manner (70% decrease for a change in Vj of +/- 75 mV). 4. These cell pairs also showed a dependency of gj on the non-junctional membrane pot...
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Experimental physiology, 1992
Cell pairs of an insect cell line (Aedes albopictus, clone C6/36) were used study simultaneously ... more Cell pairs of an insect cell line (Aedes albopictus, clone C6/36) were used study simultaneously the diffusional and electrical properties of intercellular junctions. Diffusion studies involved injection of fluorescent molecules into one cell of a cell pair and visual inspection of their intercellular redistribution. Electrical measurements involved a dual voltage clamp method and whole-cell recording with patch pipette. The voltage clamp protocol was aimed at examining the dependency of the junctional conductance, gj, on membrane potential, Vm. Cell pairs exhibiting a voltage-dependent gj were found to allow intercellular diffusion of Lucifer Yellow CH (molecular mass, 443 Da), but not of FITC-dextran (molecular mass, 4,400 Da). This response pattern is consistent with the presence of gap junctions in the intercellular junctions. Cell pairs showing no voltage dependence of gj were found to permit intercellular diffusion of both Lucifer Yellow CH and FITC-dextran (dextran labelled w...
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Papers by christoph kempf