Insights into the structure/function of hepatocyte growth factor/scatter factor from studies with individual domains.

2.50
Hdl Handle:
http://hdl.handle.net/10541/70413
Title:
Insights into the structure/function of hepatocyte growth factor/scatter factor from studies with individual domains.
Authors:
Holmes, O; Pillozzi, S; Deakin, Jon A; Carafoli, F; Kemp, L; Butler, P J G; Lyon, Malcolm; Gherardi, Ermanno
Abstract:
Hepatocyte growth factor/scatter factor (HGF/SF), the ligand for the receptor tyrosine kinase encoded by the c-Met proto-oncogene, is a multidomain protein structurally related to the pro-enzyme plasminogen and with major roles in development, tissue regeneration and cancer. We have expressed the N-terminal (N) domain, the four kringle domains (K1 to K4) and the serine proteinase homology domain (SP) of HGF/SF individually in yeast or mammalian cells and studied their ability to: (i) bind the Met receptor as well as heparan sulphate and dermatan sulphate co-receptors, (ii) activate Met in target cells and, (iii) map their binding sites onto the beta-propeller domain of Met. The N, K1 and SP domains bound Met directly with comparable affinities (K(d)=2.4, 3.3 and 1.4 microM). The same domains also bound heparin with decreasing affinities (N>K1>>SP) but only the N domain bound dermatan sulphate. Three kringle domains (K1, K2 and K4) displayed agonistic activity on target cells. In contrast, the N and SP domains, although capable of Met binding, displayed no or little activity. Further, cross-linking experiments demonstrated that both the N domain and kringles 1-2 bind the beta-chain moiety (amino acid residues 308-514) of the Met beta-propeller. In summary, the K1, K2 and K4 domains of HGF/SF are sufficient for Met activation, whereas the N and SP domains are not, although the latter domains contribute additional binding sites necessary for receptor activation by full length HGF/SF. The results provide new insights into the structure/function of HGF/SF and a basis for engineering the N and K1 domains as receptor antagonists for cancer therapy.
Affiliation:
MRC Centre, Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK.
Citation:
Insights into the structure/function of hepatocyte growth factor/scatter factor from studies with individual domains. 2007, 367 (2):395-408 J. Mol. Biol.
Journal:
Journal of Molecular Biology
Issue Date:
23-Mar-2007
URI:
http://hdl.handle.net/10541/70413
DOI:
10.1016/j.jmb.2006.12.061
PubMed ID:
17258232
Type:
Article
Language:
en
ISSN:
0022-2836
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorHolmes, O-
dc.contributor.authorPillozzi, S-
dc.contributor.authorDeakin, Jon A-
dc.contributor.authorCarafoli, F-
dc.contributor.authorKemp, L-
dc.contributor.authorButler, P J G-
dc.contributor.authorLyon, Malcolm-
dc.contributor.authorGherardi, Ermanno-
dc.date.accessioned2009-06-12T15:03:39Z-
dc.date.available2009-06-12T15:03:39Z-
dc.date.issued2007-03-23-
dc.identifier.citationInsights into the structure/function of hepatocyte growth factor/scatter factor from studies with individual domains. 2007, 367 (2):395-408 J. Mol. Biol.en
dc.identifier.issn0022-2836-
dc.identifier.pmid17258232-
dc.identifier.doi10.1016/j.jmb.2006.12.061-
dc.identifier.urihttp://hdl.handle.net/10541/70413-
dc.description.abstractHepatocyte growth factor/scatter factor (HGF/SF), the ligand for the receptor tyrosine kinase encoded by the c-Met proto-oncogene, is a multidomain protein structurally related to the pro-enzyme plasminogen and with major roles in development, tissue regeneration and cancer. We have expressed the N-terminal (N) domain, the four kringle domains (K1 to K4) and the serine proteinase homology domain (SP) of HGF/SF individually in yeast or mammalian cells and studied their ability to: (i) bind the Met receptor as well as heparan sulphate and dermatan sulphate co-receptors, (ii) activate Met in target cells and, (iii) map their binding sites onto the beta-propeller domain of Met. The N, K1 and SP domains bound Met directly with comparable affinities (K(d)=2.4, 3.3 and 1.4 microM). The same domains also bound heparin with decreasing affinities (N>K1>>SP) but only the N domain bound dermatan sulphate. Three kringle domains (K1, K2 and K4) displayed agonistic activity on target cells. In contrast, the N and SP domains, although capable of Met binding, displayed no or little activity. Further, cross-linking experiments demonstrated that both the N domain and kringles 1-2 bind the beta-chain moiety (amino acid residues 308-514) of the Met beta-propeller. In summary, the K1, K2 and K4 domains of HGF/SF are sufficient for Met activation, whereas the N and SP domains are not, although the latter domains contribute additional binding sites necessary for receptor activation by full length HGF/SF. The results provide new insights into the structure/function of HGF/SF and a basis for engineering the N and K1 domains as receptor antagonists for cancer therapy.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshBinding Sites-
dc.subject.meshCell Line-
dc.subject.meshCell Movement-
dc.subject.meshCricetinae-
dc.subject.meshCricetulus-
dc.subject.meshDermatan Sulfate-
dc.subject.meshDogs-
dc.subject.meshElectrophoretic Mobility Shift Assay-
dc.subject.meshEnzyme Activation-
dc.subject.meshExtracellular Signal-Regulated MAP Kinases-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshHepatocyte Growth Factor-
dc.subject.meshHumans-
dc.subject.meshKringles-
dc.subject.meshMice-
dc.subject.meshMutation-
dc.subject.meshPhosphorylation-
dc.subject.meshPichia-
dc.subject.meshProtein Binding-
dc.subject.meshProtein Structure, Tertiary-
dc.subject.meshProto-Oncogene Proteins c-met-
dc.subject.meshSerine Endopeptidases-
dc.subject.meshStructure-Activity Relationship-
dc.titleInsights into the structure/function of hepatocyte growth factor/scatter factor from studies with individual domains.en
dc.typeArticleen
dc.contributor.departmentMRC Centre, Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK.en
dc.identifier.journalJournal of Molecular Biologyen

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