NR AUNG
AU Cuccioloni,M.; Amici,M.; Eleuteri,A.M.; Biagetti,M.; Barocci,S.; Angeletti,M.
TI Binding of recombinant PrPc to human plasminogen: kinetic and thermodynamic study using a resonant mirror biosensor.
QU Proteins 2005 Feb 15; 58(3): 728-34
PT journal article
AB Transmissible spongiform encephalopathies are a class of sporadic, genetic and transmissible neurodegenerative diseases that affect both humans and animals. Propagation of these diseases is thought to be due to the misfolding of a neuronal glyco-protein, PrPc, into a pathological insoluble conformer, PrPsc. In earlier works, some serum components were identified as exclusive PrPsc-interacting proteins (Fisher et al., Nature 2000;408:479), and thus those macromolecules were thought to represent a potential diagnostic endogenous factor discriminating between normal and pathological prion proteins. In contrast, in agreement with a recent work (Kornblatt et al., Biochem Biophys Res Commun 2003;305:518), in this paper we present a detailed thermodynamic and kinetic characterization of the interaction between recombinant bovine PrP(c 25-242) and the human serum component plasminogen, measured using a resonant mirror technique: our results reveal a high-affinity interaction between the two binding partners. For comparison, the complex obtained from the purified full-length PrPc and human plasminogen was also studied: both prion proteins (the recombinant bovine PrP(c 25-242) and the purified full-length PrPc) are able to bind human plasminogen. Both kinetic and thermodynamic parameters are affected by the modulation exerted by the H(+) ions in solution. Moreover, the analysis of binding, according to canonical linkage relationships, suggests the involvement of a His residue, consistent with the interaction between other serine (pro)enzymes and their ligands.
MH Animals; *Biosensing Techniques; Cattle; Chromatography; Glycosylation; Histidine/chemistry; Humans; Hydrogen/chemistry; Hydrogen-Ion Concentration; Ions; Kinetics; Ligands; Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase/chemistry; Plasminogen/*chemistry; PrPc Proteins/*chemistry; PrPsc Proteins/*chemistry; Prions/chemistry; Protein Binding; Protein Isoforms; Proteins/chemistry; Proteomics/*methods; Protons; Recombinant Proteins/*chemistry; Research Support, Non-U.S. Gov't; Structure-Activity Relationship; Thermodynamics
AD
MCAB Department, University of Camerino, Italy. massimiliano.cuccioloni@unicam.it
SP englisch
PO USA