title: Allosteric antibody inhibition of human hepsin protease.
abstract: Hepsin is a type II transmembrane serine protease that is expressed in several human tissues. Overexpression of hepsin has been found to correlate with tumour progression and metastasis, which is so far best studied for prostate cancer, where more than 90% of such tumours show this characteristic. To enable improved future patient treatment, we have developed a monoclonal humanized antibody that selectively inhibits human hepsin and does not inhibit other related proteases. We found that our antibody, hH35, potently inhibits hepsin enzymatic activity at nanomolar concentrations. Kinetic characterization revealed non-linear, slow, tight-binding inhibition. This correlates with the crystal structure we obtained for the human hepsin-hH35 antibody Fab fragment complex, which showed that the antibody binds hepsin around α3-helix, located far from the active centre. The unique allosteric mode of inhibition of hH35 is distinct from the recently described HGFA (hepatocyte growth factor activator) allosteric antibody inhibition. We further explain how a small change in the antibody design induces dramatic structural rearrangements in the hepsin antigen upon binding, leading to complete enzyme inactivation.
authors: Gene Center Munich, Department of Biochemistry, Ludwig-Maximilians-Universität München, Munich, Germany.
authors: Tobias
authors: T
authors: Koschubs
authors: Stefan
authors: S
authors: Dengl
authors: Harald
authors: H
authors: Dürr
authors: Klaus
authors: K
authors: Kaluza
authors: Guy
authors: G
authors: Georges
authors: Christiane
authors: C
authors: Hartl
authors: Stefan
authors: S
authors: Jennewein
authors: Martin
authors: M
authors: Lanzendörfer
authors: Johannes
authors: J
authors: Auer
authors: Alvin
authors: A
authors: Stern
authors: Kuo-Sen
authors: KS
authors: Huang
authors: Kathryn
authors: K
authors: Packman
authors: Ueli
authors: U
authors: Gubler
authors: Dirk
authors: D
authors: Kostrewa
authors: Stefan
authors: S
authors: Ries
authors: Silke
authors: S
authors: Hansen
authors: Ulrich
authors: U
authors: Kohnert
authors: Patrick
authors: P
authors: Cramer
authors: Olaf
authors: O
authors: Mundigl
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology*; D018360:Crystallography, X-Ray; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D011487:Protein Conformation; D012697:Serine Endopeptidases / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D015842:Serine Proteinase Inhibitors; C081092:HGF activator; D012697:Serine Endopeptidases; C055645:hepsin