Influence of the Stability of a Fused Protein and Its Distance to the Amyloidogenic Segment on Fibril Formation

Conversion of native proteins into amyloid fibrils is irreversible and therefore it is difficult to study the interdependence of conformational stability and fibrillation by thermodynamic analyses. Here we approached this problem by fusing amyloidogenic poly-alanine segments derived from the N-terminal domain of the nuclear poly (A) binding protein PABPN1 with a well studied, reversibly unfolding protein, CspB from Bacillus subtilis. Earlier studies had indicated that CspB could maintain its folded structure in fibrils, when it was separated from the amyloidogenic segment by a long linker. When CspB is directly fused with the amyloidogenic segment, it unfolds because its N-terminal chain region becomes integrated into the fibrillar core, as shown by protease mapping experiments. Spacers of either 3 or 16 residues between CspB and the amyloidogenic segment were not sufficient to prevent this loss of CspB structure. Since the low thermodynamic stability of CspB (ΔGD = 12.4 kJ/mol) might be responsible for unfolding and integration of CspB into fibrils, fusions with a CspB mutant with enhanced thermodynamic stability (ΔGD = 26.9 kJ/mol) were studied. This strongly stabilized CspB remained folded and prevented fibril formation in all fusions. Our data show that the conformational stability of a linked, independently structured protein domain can control fibril formation

Masses - Don Mus.Ms. 1740 : V (X), org; B|b

Die alte Signatur mit BleistiftFranz Xaver SchmidQuelle: manuscript. - Provenienz: Fürstlich Fürstenbergische Hofbibliothek, DonaueschingenDeutsche Messe. | von | Franz Xaver Schmi

Dissertatio Ivridica Inavgvralis De Hypothecarvm Legalivm Praerogativa

Altdorf, Univ., Jur. Diss., 1773Qvam ... In Academia Altorfina ... Disceptationi Pvblicae Die XXX. Mart. A. S. R. MDCCLXXIII. Proposvit M. Franciscvs Xaverius Schmid Raittenbvchensis Franco.Autopsie nach Ex. der ULB Sachsen-AnhaltVorlageform des Erscheinungsvermerks: Altorfii Typis Ioannis Pavli Meyeri

Der neue Faust / von Tertullian Faber

DER NEUE FAUST / VON TERTULLIAN FABER Der neue Faust / von Tertullian Faber (1) Einband (3) Titelseite (5) Zueignung (6) Das alte Haus (11) Ungewitter (23) Sonnenschein (37

Studien zu einem neuen Faust / Von Dr. Xaver Schmid

STUDIEN ZU EINEM NEUEN FAUST / VON DR. XAVER SCHMID Studien zu einem neuen Faust / Von Dr. Xaver Schmid (1) Einband (1) Titelseite (5) Kapitel (7) Zueignung (9) Das alte Haus (19) Ungewitter (43) Sonnenschein (71

Der neue Faust / von Tertullian Faber

DER NEUE FAUST / VON TERTULLIAN FABER Der neue Faust / von Tertullian Faber (1) Einband (1) Titelseite (3) Titelseite (5) Zueignung (7) Das alte Haus (17) Ungewitter (41) Sonnenschein (69

Wie überlebt das Christentum?

Kaufmann F-X. Wie überlebt das Christentum? In: Schmid H, ed. Angebot der Volkskirchen und Nachfrage des Kirchenvolks. Leiten, Lenken, Gestalten. Vol 29. Wien: Lit; 2009: 93-103

A Kinetic Approach to Determining the Conformational Stability of a Protein That Dimerizes after Folding

A strongly stabilized form of the β1 domain of the streptococcal protein G, termed Gβ1-M2, was previously obtained by an in vitro selection method for stabilized protein variants. It contains four substitutions, but how they contribute to the Gibbs free energy of denaturation (Δ<i>G</i>_D) could not be determined, because, unlike the wild-type protein, Gβ1-M2 dimerizes in a spectroscopically silent reaction. Here we determined the Δ<i>G</i>_D of the folded Gβ1-M2 monomer by using a kinetic approach that uncouples the folding of the monomer from dimerization. The conformational equilibration of the monomer is faster than dimer formation, and therefore, its stability constant could be determined from the ratio of the rate constants for monomer unfolding and refolding. In this approach, double-mixing experiments were essential for uncovering the unfolding kinetics of the transient Gβ1-M2 monomer and the association of the monomers after their folding. The analysis revealed that the selected substitutions stabilize the Gβ1-M2 monomer by 15 kJ mol^–1 in an additive fashion. The combination of single- and double-mixing kinetic experiments thus allowed us to determine the thermodynamic stability of a transient species that is inaccessible in equilibrium experiments. It can be applied for proteins in which monomer folding and oligomerization are kinetically uncoupled

Wohlfahrt, Arbeit und Staat unter den Bedingungen von Globalisierung und Individualisierung

Kaufmann F-X. Wohlfahrt, Arbeit und Staat unter den Bedingungen von Globalisierung und Individualisierung. In: Schmid H, Walter-Busch E, Geiser T, eds. Arbeit in der Schweiz des 20. Jahrhunderts: Wirtschaftliche, rechtliche und soziale Perspektiven. Bern: Haupt; 1998: 1-26