Structural Basis for the Binding Specificity of Human Recepteur d'Origine Nantais (RON) Receptor Tyrosine Kinase to Macrophage-stimulating Protein

Chao, Kinlin; Gorlatova, Natalia; Eisenstein, Edward; Herzberg, Osnat

doi:10.1074/jbc.m114.594341

Cited by 18 publications

(23 citation statements)

References 72 publications

(50 reference statements)

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“…In addition to NK1, there is another Met-binding interface in the SP domain identified by a crystallographic analysis of a complex between the HGF SP and a fragment of Met comprising Sema and PSI domains 11 . The functional importance of this interface is validated by a mutagenesis study 12 , as well as by the fact that the same interface was found in the crystal structure of complex between the HGF homologue MSP and the Met homologue Ron 13 . Although HGF SP alone cannot activate Met 12 , a simultaneous addition of two HGF fragments, one encompassing N to K4 (called NK4 hereafter) and the other comprising SP, cooperatively activates Met 14 , indicating that the two binding segments can be reconstituted into an active ligand in the absence of the chain-connecting disufide bridge.…”

mentioning

confidence: 77%

Probing conformational and functional states of human hepatocyte growth factor by a panel of monoclonal antibodies

Sakai

Ogasawara

Kaneko

et al. 2016

Sci Rep

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HGF-Met signaling contributes to various biological events by controlling cell migration. Since the abnormal activation of Met receptor causes cancer progression, inhibitors such as neutralizing antibodies are regarded as promising therapeutics. HGF is secreted as a single-chain (sc) precursor and is processed by extracellular proteases to generate disulfide-bonded two-chain (tc) HGF. Although this proteolytic processing of HGF is necessary for its biological activity, exactly how the proteolysis leads to the conversion of HGF to the active form is still unclar due to the lack of structural information. In order to gain insights about this point, we generated 6 antibodies against HGF. All antibodies recognized different epitopes on the native HGF protein and showed distinct effects when tested in a cell-based HGF-Met signaling assay. They included one antibody (t1E4) that strongly blocks Met activation by tcHGF, as well as one antibody (t8E4) exclusively recognizing the active tcHGF but not inactive scHGF. Thus, a panel of anti-HGF antibodies suitable for probing the structural mechanism of HGF activation were obtained.

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mentioning

confidence: 77%

Probing conformational and functional states of human hepatocyte growth factor by a panel of monoclonal antibodies

Sakai

Ogasawara

Kaneko

et al. 2016

Sci Rep

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“…Therefore only monomers of full length MSP are required to activate Ron and induce Ron homodimerization [38]. However, analytical ultracentrifugation and crystallographic studies reported by Chao et al revealed the presence of MSP:Ron complexes with a 2:2 stoichiometry [42]. These complexes were in relatively low abundance and had weaker binding affinity compared to the 1:1 and 1:2 complexes, but their presence suggests a possible role for MSP:MSP interaction within the Ron signaling complex although the physiological relevance is currently unclear.…”

Section: Discussionmentioning

confidence: 99%

Analogs of the hepatocyte growth factor and macrophage-stimulating protein hinge regions act as Met and Ron dual inhibitors in pancreatic cancer cells

Church

Vanderwerff²,

Riggers³

et al. 2016

Anti-Cancer Drugs

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Pancreatic cancer is among the leading causes of cancer death in the United States with limited effective treatment options. A major contributor to the formation and persistence of pancreatic cancer is the dysregulation of the hepatocyte growth factor (HGF)/Met (HGF receptor) and the macrophage stimulating protein (MSP)/Ron (MSP receptor) systems. These systems normally mediate a variety of cellular behaviors including proliferation, survival, and migration, but are often over-activated in pancreatic cancer and contribute to cancer progression. Previous studies have shown that HGF must dimerize in order to activate Met. Small molecule antagonists with homology to a “hinge” region within the putative dimerization domain of HGF have been developed that bind to HGF and block dimerization, therefore inhibiting Met signaling. Due to the structural and sequence homology between MSP and HGF, we hypothesized that the inhibition of HGF by the hinge analogs may extend to MSP. The primary purpose of this “proof-of-concept” study was to determine if hinge analogs could inhibit cellular responses to both HGF and MSP in pancreatic cancer cells. Our results demonstrated that these compounds inhibited HGF and MSP activity. Hinge analog treatment resulted in decreased Met and Ron activation, and suppressed malignant cell behaviors including proliferation, migration, and invasion in pancreatic cancer cells in vitro. These results suggest that the hinge analogs represent a novel group of molecules that may offer a therapeutic approach for the treatment of pancreatic cancer and warrant further development and optimization.

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“…The results of computational alanine scanning reported here revealed that Glu 287 , Pro 288 , Glu 289 , and His 424 of RON located at the interface of RON-MSPb complex are crucial residues for its binding to MSPb. 76 According to ΔΔG values, Pro 288 , Glu 287 , and Glu 289 of RON are the next most important amino acids after Arg 220 (see Table 3). It seems that the importance of these residues is related to their interactions with more than one residues on MSP (except for Pro 288 ).…”

Section: Resultsmentioning

confidence: 99%

Characterizing the hotspots involved in RON-MSPβ complex formation using in silico alanine scanning mutagenesis and molecular dynamics simulation

Zarei¹,

Hamzeh‐Mivehroud²,

Benvenuti³

et al. 2017

Adv Pharm Bull

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Purpose: Implication of protein-protein interactions (PPIs) in development of many diseases such as cancer makes them attractive for therapeutic intervention and rational drug design. RON (Recepteur d’Origine Nantais) tyrosine kinase receptor has gained considerable attention as promising target in cancer therapy. The activation of RON via its ligand, macrophage stimulation protein (MSP) is the most common mechanism of activation for this receptor. The aim of the current study was to perform in silico alanine scanning mutagenesis and to calculate binding energy for prediction of hot spots in protein-protein interface between RON and MSPβ chain (MSPβ). Methods: In this work the residues at the interface of RON-MSPβ complex were mutated to alanine and then molecular dynamics simulation was used to calculate binding free energy. Results: The results revealed that Gln193, Arg220, Glu287, Pro288, Glu289, and His424 residues from RON and Arg521, His528, Ser565, Glu658, and Arg683 from MSPβ may play important roles in protein-protein interaction between RON and MSP. Conclusion: Identification of these RON hot spots is important in designing anti-RON drugs when the aim is to disrupt RON-MSP interaction. In the same way, the acquired information regarding the critical amino acids of MSPβ can be used in the process of rational drug design for developing MSP antagonizing agents, the development of novel MSP mimicking peptides where inhibition of RON activation is required, and the design of experimental site directed mutagenesis studies.

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Structural Basis for the Binding Specificity of Human Recepteur d'Origine Nantais (RON) Receptor Tyrosine Kinase to Macrophage-stimulating Protein

Cited by 18 publications

References 72 publications

Probing conformational and functional states of human hepatocyte growth factor by a panel of monoclonal antibodies

Probing conformational and functional states of human hepatocyte growth factor by a panel of monoclonal antibodies

Analogs of the hepatocyte growth factor and macrophage-stimulating protein hinge regions act as Met and Ron dual inhibitors in pancreatic cancer cells

Characterizing the hotspots involved in RON-MSPβ complex formation using in silico alanine scanning mutagenesis and molecular dynamics simulation

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