TY  - JOUR
A1  - Hanisch, Uwe-Karsten
A1  - Van Rossum, Denise
A1  - Xie, Yiheng
A1  - Misselwitz, Rolf
A1  - Auriola, Seppo
A1  - Goldstein, Gundars
A1  - Koistinaho, Jari
A1  - Kettemann, Helmut
A1  - Möller, Thomas
A1  - Gast, Klaus
T1  - The microglia-activating potential of thrombin : the protease is not involved in the induction of proinflammatory cytokines and chemokines
N2  - The serine protease thrombin is known as a blood coagulation factor. Through limited cleavage of proteinase- activated receptors it can also control growth and functions in various cell types, including neurons, astrocytes, and microglia ( brain macrophages). A number of previous studies indicated that thrombin induces the release of proinflammatory cytokines and chemokines from microglial cells, suggesting another important role for the protease beyond hemostasis. In the present report, we provide evidence that this effect is not mediated by any proteolytic or non- proteolytic mechanism involving thrombin proper. Inhibition of the enzymatic thrombin activity did not affect the microglial release response. Instead the cyto-/chemokine-inducing activity solely resided in a high molecular weight protein fraction that could be isolated in trace amounts even from apparently homogenous alpha- and gamma-thrombin preparations. High molecular weight material contained thrombin-derived peptides as revealed by mass spectrometry but was devoid of thrombin-like enzymatic activity. Separated from the high molecular weight fraction by fast protein liquid chromatography, enzymatically intact alpha- and gamma-thrombin failed to trigger any release. Our findings may force a revision of the notion that thrombin itself is a direct proinflammatory release signal for microglia. In addition, they could be relevant for the study of other cellular activities and their assignment to this protease
Y1  - 2004
ER  - 
TY  - JOUR
A1  - Hanisch, Uwe-Karsten
A1  - van Rossum, D.
A1  - Gast, Klaus
A1  - Misselwitz, Rolf
A1  - Goldstein, Gundars
A1  - Koistinaho, Jari
A1  - Möller, Thomas
T1  - The microglia-activating potential of thrombin : is the protease able to induce cyto- and chemokines?
Y1  - 2004
ER  - 
TY  - JOUR
A1  - Fabian, Heinz
A1  - Gast, Klaus
A1  - Laue, Michael
A1  - Misselwitz, Rolf
A1  - Uchanska-Ziegler, Barbara
A1  - Ziegler, Andreas
A1  - Naumann, Dieter
T1  - Early stages of misfolding and association of beta2-microglobulin : insights from infrared spectroscopy and dynamic light scattering
N2  - Conformational changes associated with the assembly of recombinant ;2-microglobulin in vitro under acidic conditions were investigated using infrared spectroscopy and static and dynamic light scattering. In parallel, the morphology of the different aggregated species obtained under defined conditions was characterized by electron microscopy. The initial salt-induced aggregate form of ;2-microglobulin, composed of small oligomers (dimers to tetramers), revealed the presence of ;-strands organized in an intramolecular-like fashion. Further particle growth was accompanied by the formation of intermolecular ;-sheet structure and led to short curved forms. An increase in temperature by only 25 °C was able to disaggregate these assemblies, followed by the formation of longer filamentous structures. In contrast, a rise in temperature up to 100 °C was associated with a reorganization of the short curved forms at the level of secondary structure and the state of assembly, leading to a species with a characteristic infrared spectrum different from those of all the other aggregates observed before, suggesting a unique overall structure. The infrared spectral features of this species were nearly identical to those of ;2-microglobulin assemblies formed at low ionic strength with agitation, indicating the presence of fibrils, which was confirmed by electron microscopy. The observed spectroscopic changes suggest that the heat-triggered conversion of the short curved assemblies into fibrils involves a reorganization of the ;-strands from an antiparallel arrangement to a parallel arrangement, with the latter being characteristic of amyloid fibrils of ;2-microglobulin.
Y1  - 2008
UR  - http://pubs.acs.org/doi/abs/10.1021/bi800279y
ER  - 
TY  - JOUR
A1  - Damaschun, Gregor
A1  - Damaschun, Hilde
A1  - Gast, Klaus
A1  - Misselwitz, Rolf
A1  - Müller, Jürgen J.
A1  - Pfeil, Wolfgang
A1  - Zirwer, Dietrich
T1  - Cold denaturation-induced conformational changes in phosphoglycerate kinase from yeast
Y1  - 1993
ER  -