Characterization of the heparin/heparan sulfate binding site of the natural cytotoxicity receptor NKp46.

2.50
Hdl Handle:
http://hdl.handle.net/10541/76274
Title:
Characterization of the heparin/heparan sulfate binding site of the natural cytotoxicity receptor NKp46.
Authors:
Zilka, Alon; Landau, Guy; Hershkovitz, Oren; Bloushtain, Noga; Bar-Ilan, Ahuva; Benchetrit, Fabrice; Fima, Eyal; Van Kuppevelt, Toin H; Gallagher, John T; Elgavish, Sharona; Porgador, Angel
Abstract:
NKp46 is a member of a group of receptors collectively termed natural cytotoxicity receptors (NCRs) that are expressed by natural killer (NK) cells. NCRs are capable of mediating direct killing of tumor and virus-infected cells by NK cells. We have recently shown that NKp46 recognizes the heparan sulfate moieties of membranal heparan sulfate proteoglycans (HSPGs), thus enabling lysis of tumor cells by NK cells. In the current study, we further examined the residues in NKp46 that may be involved in heparan sulfate binding on tumor cells. On the basis of both the electrostatic potential map and comparison to the heparin binding site on human fibronectin, we predicted a continuous region containing the basic amino acids K133, R136, H139, R142, and K146 to be involved in NKp46 binding to heparan sulfate. Mutating these amino acids on NKp46D2 to noncharged amino acids retained its virus binding capacity but reduced its binding to tumor cells with a 10-100 fold lower K(D) when tested for direct binding to heparin. The minimal length of the heparin/heparan sulfate epitope recognized by NKp46 was eight saccharides as predicted from the structure and proven by testing heparin oligomers. Testing selectively monodesulfated heparin oligomers emphasized the specific contributions of O-sulfation, N-sulfation, and N-acetylation to epitope recognition by NKp46. The characterization of heparan sulfate binding region in NKp46 offers further insight into the identity of the ligands for NKp46 and the interaction of NK and cancers.
Affiliation:
Department of Microbiology and Immunology, Faculty of Health Sciences, and Cancer Research Center, Ben Gurion University of the Negev, Beer Sheva 84105, Israel.
Citation:
Characterization of the heparin/heparan sulfate binding site of the natural cytotoxicity receptor NKp46. 2005, 44 (44):14477-85 Biochemistry
Journal:
Biochemistry
Issue Date:
8-Nov-2005
URI:
http://hdl.handle.net/10541/76274
DOI:
10.1021/bi051241s
PubMed ID:
16262248
Type:
Article
Language:
en
ISSN:
0006-2960
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorZilka, Alon-
dc.contributor.authorLandau, Guy-
dc.contributor.authorHershkovitz, Oren-
dc.contributor.authorBloushtain, Noga-
dc.contributor.authorBar-Ilan, Ahuva-
dc.contributor.authorBenchetrit, Fabrice-
dc.contributor.authorFima, Eyal-
dc.contributor.authorVan Kuppevelt, Toin H-
dc.contributor.authorGallagher, John T-
dc.contributor.authorElgavish, Sharona-
dc.contributor.authorPorgador, Angel-
dc.date.accessioned2009-08-04T17:11:40Z-
dc.date.available2009-08-04T17:11:40Z-
dc.date.issued2005-11-08-
dc.identifier.citationCharacterization of the heparin/heparan sulfate binding site of the natural cytotoxicity receptor NKp46. 2005, 44 (44):14477-85 Biochemistryen
dc.identifier.issn0006-2960-
dc.identifier.pmid16262248-
dc.identifier.doi10.1021/bi051241s-
dc.identifier.urihttp://hdl.handle.net/10541/76274-
dc.description.abstractNKp46 is a member of a group of receptors collectively termed natural cytotoxicity receptors (NCRs) that are expressed by natural killer (NK) cells. NCRs are capable of mediating direct killing of tumor and virus-infected cells by NK cells. We have recently shown that NKp46 recognizes the heparan sulfate moieties of membranal heparan sulfate proteoglycans (HSPGs), thus enabling lysis of tumor cells by NK cells. In the current study, we further examined the residues in NKp46 that may be involved in heparan sulfate binding on tumor cells. On the basis of both the electrostatic potential map and comparison to the heparin binding site on human fibronectin, we predicted a continuous region containing the basic amino acids K133, R136, H139, R142, and K146 to be involved in NKp46 binding to heparan sulfate. Mutating these amino acids on NKp46D2 to noncharged amino acids retained its virus binding capacity but reduced its binding to tumor cells with a 10-100 fold lower K(D) when tested for direct binding to heparin. The minimal length of the heparin/heparan sulfate epitope recognized by NKp46 was eight saccharides as predicted from the structure and proven by testing heparin oligomers. Testing selectively monodesulfated heparin oligomers emphasized the specific contributions of O-sulfation, N-sulfation, and N-acetylation to epitope recognition by NKp46. The characterization of heparan sulfate binding region in NKp46 offers further insight into the identity of the ligands for NKp46 and the interaction of NK and cancers.en
dc.language.isoenen
dc.subjectCell Line Tumouren
dc.subject.meshAmino Acid Sequence-
dc.subject.meshAnticoagulants-
dc.subject.meshBinding Sites-
dc.subject.meshCell Line, Tumor-
dc.subject.meshHeparin-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshHumans-
dc.subject.meshMembrane Glycoproteins-
dc.subject.meshModels, Molecular-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshMutation-
dc.subject.meshNatural Cytotoxicity Triggering Receptor 1-
dc.subject.meshProtein Binding-
dc.subject.meshProtein Structure, Secondary-
dc.subject.meshProtein Structure, Tertiary-
dc.subject.meshReceptors, Immunologic-
dc.subject.meshSequence Alignment-
dc.titleCharacterization of the heparin/heparan sulfate binding site of the natural cytotoxicity receptor NKp46.en
dc.typeArticleen
dc.contributor.departmentDepartment of Microbiology and Immunology, Faculty of Health Sciences, and Cancer Research Center, Ben Gurion University of the Negev, Beer Sheva 84105, Israel.en
dc.identifier.journalBiochemistryen

Related articles on PubMed

All Items in Christie are protected by copyright, with all rights reserved, unless otherwise indicated.