Structural and Functional Basis of the Serine Protease-like Hepatocyte Growth Factor β-Chain in Met Binding and Signaling

Kirchhofer, Daniel; Yao, Xiaoyi; Peek, Mark; Eigenbrot, Charles; Lipari, Michael T.; Billeci, Karen L.; Maun, Henry R.; Moran, Paul; Santell, Lydia; Wiesmann, Christian; Lazarus, Robert A.

doi:10.1074/jbc.m404795200

Cited by 83 publications

(118 citation statements)

References 52 publications

(77 reference statements)

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“…2 J, which converge to a common structure for the nk3 segment and differ by a translation of the sp and, to a lesser extent, the k4 domain. Consequently, the receptor binding sites of the k1 (30) and sp (21,22) domains, shown as blue and red patches in Fig. 2 (22) or 3D models (15) of the MET sema, cr, and ig1 domains could be docked accurately into the EM density maps (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…5F) support this interpretation. The nk4 fragment (39) or single-chain HGF͞SF may thus fail to yield a 2:2 complex with MET because in nk4 the sp domain is missing and in single-chain HGF͞SF the domain is locked in a conformation that only enables low affinity binding (21); lack of binding through the sp domain may destabilize dimerization via the n and k1 domains. As for the proposed role for the sp domain in MET dimerization (22), the results of our study suggest an alternative one, namely in the formation of higher-order signaling complexes.…”

Section: Discussionmentioning

confidence: 99%

“…Several nuclear magnetic resonance and crystal structures of HGF͞SF fragments corresponding to the n domain (17), the n and k1 domains (nk1) (18)(19)(20), and the sp domain (21), and a crystal structure of the sema and cr domains of MET in complex with the sp domain of HGF͞SF (22) have provided important insights but failed to show the interdomain interactions and the relevant changes which underlie biological activity.…”

Section: C-e)mentioning

confidence: 99%

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Structural basis of hepatocyte growth factor/scatter factor and MET signalling

Gherardi

Sandin

Petoukhov

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

199

194

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The polypeptide growth factor, hepatocyte growth factor͞scatter factor (HGF͞SF), shares the multidomain structure and proteolytic mechanism of activation of plasminogen and other complex serine proteinases. HGF͞SF, however, has no enzymatic activity. Instead, it controls the growth, morphogenesis, or migration of epithelial, endothelial, and muscle progenitor cells through the receptor tyrosine kinase MET. Using small-angle x-ray scattering and cryoelectron microscopy, we show that conversion of pro(single-chain)-HGF͞SF into the active two-chain form is associated with a major structural transition from a compact, closed conformation to an elongated, open one. We also report the structure of a complex between two-chain HGF͞SF and the MET ectodomain (MET928) with 1:1 stoichiometry in which the N-terminal and first kringle domain of HGF͞SF contact the face of the seven-blade ␤-propeller domain of MET harboring the loops connecting the ␤-strands b-c and d-a, whereas the C-terminal serine proteinase homology domain binds the opposite ''b'' face. Finally, we describe a complex with 2:2 stoichiometry between two-chain HGF͞SF and a truncated form of the MET ectodomain (MET567), which is assembled around the dimerization interface seen in the crystal structure of the NK1 fragment of HGF͞SF and displays the features of a functional, signaling unit. The study shows how the proteolytic mechanism of activation of the complex proteinases has been adapted to cell signaling in vertebrate organisms, offers a description of monomeric and dimeric ligand-receptor complexes, and provides a foundation to the structural basis of HGF͞SF-MET signaling.cell signaling ͉ plasminogen ͉ serine proteinases ͉ kringle ͉ x-ray scattering H epatocyte growth factor͞scatter factor (HGF͞SF) (1-6) are vertebrate-specific polypeptide growth factors with a domain structure related to that of plasminogen (7). Interest in HGF͞SF and its receptor MET (8) stems from unique biological roles in embryogenesis (9-11), tissue regeneration (12, 13), and cancer (14). These activities have led to a strong interest in the structure of the molecules as this knowledge may underpin the development of MET-based therapeutics.HGF͞SF consists of six domains: an N-terminal domain (n), four copies of the kringle domain (k1-k4), and a C-terminal domain (sp) structurally related to the catalytic domain of serine proteinases (Fig. 1A). The factor is synthesized as a precursor protein (pro-or single-chain HGF͞SF) and is proteolytically processed to a two-chain form by cleavage of the linker connecting the k4 and sp domains ( Fig. 1 A and B). Single-chain HGF͞SF binds MET (15, 16) but is unable to induce biological responses, for example, dispersion of MDCK cell colonies, even at concentrations 100-fold higher than two-chain HGF͞SF (Fig. 1 C-E).MET is also synthesized as a single-chain precursor that is cleaved by furin yielding an N-terminal ␣-chain and a C-terminal ␤-chain. The MET ectodomain consists of two moieties: the large, N-terminal sema domain, which is responsible f...

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: C-e)mentioning

confidence: 99%

See 1 more Smart Citation

Structural basis of hepatocyte growth factor/scatter factor and MET signalling

Gherardi

Sandin

Petoukhov

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

199

194

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“…By examining the MSP crystal structure data (PDB ID: 2ASU) (43), we note that R689 is located on the surface of the MSP β-chain and within a positively charged patch (L13 loop) that consists of a cluster of arginine residues including R683, which has been shown to be essential for MST1R binding (44) (Figure 3). The L13 loop borders the S1 specificity pocket (involving Y661, Q522 and Q568) and together, form the putative high-affinity binding surface for MST1R, resembling the structurally determined interaction between hepatocyte growth factor (scatter factor), which is structurally related to MSP, and its receptor, MET (45,46). Furthermore, the proximity of R689C to a conserved cysteine residue (C685) that establishes a critical disulfide bond with C657 may introduce aberrant bond formations and disrupt the structural integrity of the adjacent receptor binding surface.…”

Section: The Associated R689c Variant and Mst1 Functionmentioning

confidence: 99%

Gene-centric association mapping of chromosome 3p implicates MST1 in IBD pathogenesis

Goyette

Lefèbvre

et al. 2008

Mucosal Immunology

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Association mapping and candidate gene studies within IBD linkage regions, as well as genomewide association studies in CD have led to the discovery of multiple risk genes, but these only Corresponding author: John D. Rioux, Montreal Heart Institute, 5000 Belanger Street, Montreal, Quebec, Canada, H1T 1C8, rioux@inflammgen.org. HHS Public Access Author Manuscript Author ManuscriptAuthor ManuscriptAuthor Manuscript explain a fraction of the genetic susceptibility observed in IBD. We have thus been pursuing a region on chromosome 3p21-22 showing linkage to CD and UC using a gene-centric association mapping approach. We identified twelve functional candidate genes by searching for literature cocitations with relevant keywords and for gene expression patterns consistent with immune/ intestinal function. We then performed an association study composed of a screening phase, where tagging SNPs were evaluated in 1020 IBD patients, and an independent replication phase in 745 IBD patients. These analyses identified and replicated significant association to IBD for four SNPs within a 1.2 Mb LD region. We then identified a non-synonymous coding variant (rs3197999, R689C) in the macrophage stimulating 1 (MST1) gene (p-value 3.62×10−6) that accounts for the association signal, and shows association to both CD and UC. MST1 encodes MSP, a protein regulating the innate immune responses to bacterial ligands. R689C is predicted to interfere with MSP binding to its receptor, suggesting a role for this gene in the pathogenesis of IBD.

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“…The first 519 amino acids of Met form a 7-bladed β-propeller domain, called sema domain, with homology to the semaphorin and plexin protein families (4,5). Following the sema domain are a cysteine-rich domain and four immunoglobulin-like domains.…”

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confidence: 99%

Structural basis for agonism and antagonism of hepatocyte growth factor

Tolbert¹,

Daugherty-Holtrop²,

Gherardi

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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Hepatocyte growth factor (HGF) is an activating ligand of the Met receptor tyrosine kinase, whose activity is essential for normal tissue development and organ regeneration but abnormal activation of Met has been implicated in growth, invasion, and metastasis of many types of solid tumors. HGF has two natural splice variants, NK1 and NK2, which contain the N-terminal domain (N) and the first kringle (K1) or the first two kringle domains of HGF. NK1, which is a Met agonist, forms a head-to-tail dimer complex in crystal structures and mutations in the NK1 dimer interface convert NK1 to a Met antagonist. In contrast, NK2 is a Met antagonist, capable of inhibiting HGF's activity in cell proliferation without clear mechanism. Here we report the crystal structure of NK2, which forms a "closed" monomeric conformation through interdomain interactions between the N-domain and the second kringle domain (K2). Mutations that were designed to open up the NK2 closed conformation by disrupting the N/K2 interface convert NK2 from a Met antagonist to an agonist. Remarkably, this mutated NK2 agonist can be converted back to an antagonist by a mutation that disrupts the NK1/NK1 dimer interface. These results reveal the molecular determinants that regulate the agonist/antagonist properties of HGF NK2 and provide critical insights into the dimerization mechanism that regulates the Met receptor activation by HGF.cancer metastasis | crystal structure | HGF agonist | HGF antagonist | Met receptor tyrosine kinase H epatocyte growth factor (HGF) and the Met tyrosine kinase receptor form a unique ligand-receptor signaling system where HGF is the only known endogenous ligand that activates Met. HGF is also known as a scattering factor, and its signaling through Met activation is involved in promoting cell proliferation, survival, migration, and branching in a variety of cell types with important roles in angiogenesis, wound repair, and cell migration during development (1, 2). Inappropriate expression of Met or HGF has been implicated in many cancers and is associated with an aggressive phenotype and poor prognosis (1, 3) (www.vai.org/met). Thus, inhibition of Met activity, either by small molecule kinase inhibitors or by protein-based HGF antagonists, has become an important and rational strategy for developing anticancer therapeutics.The Met extracellular domain (ECD), which encompasses approximately 900 amino acids (25-932), is cleaved into α and β chains by the furin protease between residues 307 and 308. The first 519 amino acids of Met form a 7-bladed β-propeller domain, called sema domain, with homology to the semaphorin and plexin protein families (4, 5). Following the sema domain are a cysteine-rich domain and four immunoglobulin-like domains. The sema domain of Met is necessary and sufficient for binding and activation of the receptor by HGF (6). It is believed that HGF induced Met activation is mediated through a formation of a 2∶2 complex where Met dimerization is primarily mediated by dimer formation of HGF (7,8).HGF is a disu...

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Structural and Functional Basis of the Serine Protease-like Hepatocyte Growth Factor β-Chain in Met Binding and Signaling

Cited by 83 publications

References 52 publications

Structural basis of hepatocyte growth factor/scatter factor and MET signalling

Structural basis of hepatocyte growth factor/scatter factor and MET signalling

Gene-centric association mapping of chromosome 3p implicates MST1 in IBD pathogenesis

Structural basis for agonism and antagonism of hepatocyte growth factor

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