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dc.contributor.authorSiddiqui, Shoib
dc.contributor.authorSchwarz, Flavio
dc.contributor.authorSpringer, Stevan
dc.contributor.authorKhedri, Zahra
dc.contributor.authorYu, Hai
dc.contributor.authorDeng, Lingquan
dc.contributor.authorVerhagen, Andrea
dc.contributor.authorNaito-Matsui, Yuko
dc.contributor.authorJiang, Weiping
dc.contributor.authorKim, Daniel
dc.contributor.authorZhou, Jie
dc.contributor.authorDing, Beibei
dc.contributor.authorChen, Xi
dc.contributor.authorVarki, Nissi
dc.contributor.authorVarki, Ajit
dc.identifier.citationSiddiqui , S , Schwarz , F , Springer , S , Khedri , Z , Yu , H , Deng , L , Verhagen , A , Naito-Matsui , Y , Jiang , W , Kim , D , Zhou , J , Ding , B , Chen , X , Varki , N & Varki , A 2017 , ' Studies on the Detection, Expression, Glycosylation, Dimerization, and Ligand Binding Properties of Mouse Siglec-E ' , The Journal of biological chemistry , vol. 292 , no. 3 , pp. 1029-1037 .
dc.identifier.otherPURE: 24643543
dc.identifier.otherPURE UUID: 24a42dc4-9cb5-4ce3-8709-094f4700113b
dc.identifier.otherPubMed: 27920204
dc.identifier.otherPubMedCentral: PMC5247637
dc.identifier.otherScopus: 85010032135
dc.description© 2017 by The American Society for Biochemistry and Molecular Biology, Inc. Published in the U.S.A. This is an Open Access article under the CC BY license (
dc.description.abstractCD33-related Siglecs are a family of proteins widely expressed on innate immune cells. Binding of sialylated glycans or other ligands triggers signals that inhibit or activate inflammation. Immunomodulation by Siglecs has been extensively studied, but relationships between structure and functions are poorly explored. Here we present new data relating to the structure and function of Siglec-E, the major CD33-related Siglec expressed on mouse neutrophils, monocytes, macrophages, and dendritic cells. We generated nine new rat monoclonal antibodies specific to mouse Siglec-E, with no cross-reactivity to Siglec-F. Although all antibodies detected Siglec-E on transfected human HEK-293T cells, only two reacted with mouse bone marrow neutrophils by flow cytometry and on spleen sections by immunohistochemistry. Moreover, whereas all antibodies recognized Siglec-E-Fc on immunoblots, binding was dependent on intact disulfide bonds and N-glycans, and only two antibodies recognized native Siglec-E within spleen lysates. Thus, we further investigated the impact of Siglec-E homodimerization. Homology-based structural modeling predicted a cysteine residue (Cys-298) in position to form a disulfide bridge between two Siglec-E polypeptides. Mutagenesis of Cys-298 confirmed its role in dimerization. In keeping with the high level of 9-O-acetylation found in mice, sialoglycan array studies indicate that this modification has complex effects on recognition by Siglec-E, in relationship to the underlying structures. However, we found no differences in phosphorylation or SHP-1 recruitment between dimeric and monomeric Siglec-E expressed on HEK293A cells. Phylogenomic analyses predicted that only some human and mouse Siglecs form disulfide-linked dimers. Notably, Siglec-9, the functionally equivalent human paralog of Siglec-E, occurs as a monomer.en
dc.relation.ispartofThe Journal of biological chemistry
dc.subjectAmino Acid Substitution
dc.subjectAntigens, CD/chemistry
dc.subjectAntigens, Differentiation, B-Lymphocyte/chemistry
dc.subjectDendritic Cells/cytology
dc.subjectGene Expression Regulation/physiology
dc.subjectMice, Knockout
dc.subjectMutation, Missense
dc.subjectProtein Multimerization/physiology
dc.subjectProtein Tyrosine Phosphatase, Non-Receptor Type 6
dc.subjectRats, Inbred Lew
dc.subjectSialic Acid Binding Immunoglobulin-like Lectins/chemistry
dc.titleStudies on the Detection, Expression, Glycosylation, Dimerization, and Ligand Binding Properties of Mouse Siglec-Een
dc.contributor.institutionSchool of Life and Medical Sciences
dc.description.statusPeer reviewed
rioxxterms.typeJournal Article/Review

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