The structure of a furin-antibody complex explains non-competitive inhibition by steric exclusion of substrate conformers.
Proprotein Convertases (PCs) represent highly selective serine proteases that activate their substrates upon proteolytic cleavage. Their inhibition is a promising strategy for the treatment of cancer and infectious diseases. Inhibitory camelid antibodies were developed, targeting the prototypical PC furin. Kinetic analyses of them revealed an enigmatic non-competitive mechanism, affecting the inhibition of large proprotein-like but not small peptidic substrates. Here we present the crystal structures of furin in complex with the antibody Nb14 and of free Nb14 at resolutions of 2.0 Å and 2.3 Å, respectively. Nb14 binds at a site distant to the substrate binding pocket to the P-domain of furin. Interestingly, no major conformational changes were observed upon complex formation, neither for the protease nor for the antibody. Inhibition of furin by Nb14 is instead explained by steric exclusion of specific substrate conformers, explaining why Nb14 inhibits the processing of bulky protein substrates but not of small peptide substrates. This mode of action was further supported by modelling studies with the ternary factor X-furin-antibody complex and a mutation that disrupted the interaction interface between furin and the antibody. The observed binding mode of Nb14 suggests a novel approach for the development of highly specific antibody-based proprotein convertase inhibitors.
Protein Crystallography Group, Leibniz Institute on Aging-Fritz Lipmann Institute (FLI), Beutenbergstr. 11, 07745 Jena, Germany.|Department of Molecular Biology, University of Salzburg, Billrothstrasse 11, A-5020 Salzburg, Austria.
Sven O
SO
Dahms
Department of Human Genetics, KU Leuven, Herestraat 49, B-3000 Leuven, Belgium.
John W M
JW
Creemers
Protein Crystallography Group, Leibniz Institute on Aging-Fritz Lipmann Institute (FLI), Beutenbergstr. 11, 07745 Jena, Germany.
Yvonne
Y
Schaub
European Molecular Biology Laboratory, Hamburg, Germany.
Gleb P
GP
Bourenkov
Department of Molecular Biology, University of Salzburg, Billrothstrasse 11, A-5020 Salzburg, Austria.
Thomas
T
Zögg
Department of Molecular Biology, University of Salzburg, Billrothstrasse 11, A-5020 Salzburg, Austria.
Hans
H
Brandstetter
Protein Crystallography Group, Leibniz Institute on Aging-Fritz Lipmann Institute (FLI), Beutenbergstr. 11, 07745 Jena, Germany.
Manuel E
ME
Than
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