Quantitative Characterization of E-selectin Interaction with Native CD44 and P-selectin Glycoprotein Ligand-1 (PSGL-1) Using a Real Time Immunoprecipitation-based Binding Assay

Handle URI:
http://hdl.handle.net/10754/594199
Title:
Quantitative Characterization of E-selectin Interaction with Native CD44 and P-selectin Glycoprotein Ligand-1 (PSGL-1) Using a Real Time Immunoprecipitation-based Binding Assay
Authors:
Abu Samra, Dina Bashir Kamil ( 0000-0002-0974-5362 ) ; Al Kilani, Alia ( 0000-0002-6428-1185 ) ; Hamdan, Samir ( 0000-0001-5192-1852 ) ; Sakashita, Kosuke ( 0000-0002-7118-5511 ) ; Gadhoum, Samah Z.; Merzaban, Jasmeen S. ( 0000-0002-7276-2907 )
Abstract:
Selectins (E-, P-, and L-selectins) interact with glycoprotein ligands to mediate the essential tethering/rolling step in cell transport and delivery that captures migrating cells from the circulating flow. In this work, we developed a real time immunoprecipitation assay on a surface plasmon resonance chip that captures native glycoforms of two well known E-selectin ligands (CD44/hematopoietic cell E-/L-selectin ligand and P-selectin glycoprotein ligand-1) from hematopoietic cell extracts. Here we present a comprehensive characterization of their binding to E-selectin. We show that both ligands bind recombinant monomeric E-selectin transiently with fast on- and fast off-rates, whereas they bind dimeric E-selectin with remarkably slow onand off-rates. This binding requires the sialyl Lewis x sugar moiety to be placed on both O- and N-glycans, and its association, but not dissociation, is sensitive to the salt concentration. Our results suggest a mechanism through which monomeric selectins mediate initial fast on and fast off kinetics to help capture cells out of the circulating shear flow; subsequently, tight binding by dimeric/oligomeric selectins is enabled to significantly slow rolling. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
KAUST Department:
Biological and Environmental Sciences and Engineering (BESE) Division
Citation:
AbuSamra DB, Al-Kilani A, Hamdan SM, Sakashita K, Gadhoum SZ, et al. (2015) Quantitative Characterization of E-selectin Interaction with Native CD44 and P-selectin Glycoprotein Ligand-1 (PSGL-1) Using a Real Time Immunoprecipitation-based Binding Assay. J Biol Chem 290: 21213–21230. Available: http://dx.doi.org/10.1074/jbc.m114.629451.
Publisher:
American Society for Biochemistry & Molecular Biology (ASBMB)
Journal:
Journal of Biological Chemistry
Issue Date:
29-Jun-2015
DOI:
10.1074/jbc.m114.629451
PubMed ID:
26124272
PubMed Central ID:
PMC4571854
Type:
Article
ISSN:
0021-9258; 1083-351X
Sponsors:
KAUST, King Abdullah University of Science and Technology
Appears in Collections:
Articles; Biological and Environmental Sciences and Engineering (BESE) Division

Full metadata record

DC FieldValue Language
dc.contributor.authorAbu Samra, Dina Bashir Kamilen
dc.contributor.authorAl Kilani, Aliaen
dc.contributor.authorHamdan, Samiren
dc.contributor.authorSakashita, Kosukeen
dc.contributor.authorGadhoum, Samah Z.en
dc.contributor.authorMerzaban, Jasmeen S.en
dc.date.accessioned2016-01-19T13:23:41Zen
dc.date.available2016-01-19T13:23:41Zen
dc.date.issued2015-06-29en
dc.identifier.citationAbuSamra DB, Al-Kilani A, Hamdan SM, Sakashita K, Gadhoum SZ, et al. (2015) Quantitative Characterization of E-selectin Interaction with Native CD44 and P-selectin Glycoprotein Ligand-1 (PSGL-1) Using a Real Time Immunoprecipitation-based Binding Assay. J Biol Chem 290: 21213–21230. Available: http://dx.doi.org/10.1074/jbc.m114.629451.en
dc.identifier.issn0021-9258en
dc.identifier.issn1083-351Xen
dc.identifier.pmid26124272en
dc.identifier.doi10.1074/jbc.m114.629451en
dc.identifier.urihttp://hdl.handle.net/10754/594199en
dc.description.abstractSelectins (E-, P-, and L-selectins) interact with glycoprotein ligands to mediate the essential tethering/rolling step in cell transport and delivery that captures migrating cells from the circulating flow. In this work, we developed a real time immunoprecipitation assay on a surface plasmon resonance chip that captures native glycoforms of two well known E-selectin ligands (CD44/hematopoietic cell E-/L-selectin ligand and P-selectin glycoprotein ligand-1) from hematopoietic cell extracts. Here we present a comprehensive characterization of their binding to E-selectin. We show that both ligands bind recombinant monomeric E-selectin transiently with fast on- and fast off-rates, whereas they bind dimeric E-selectin with remarkably slow onand off-rates. This binding requires the sialyl Lewis x sugar moiety to be placed on both O- and N-glycans, and its association, but not dissociation, is sensitive to the salt concentration. Our results suggest a mechanism through which monomeric selectins mediate initial fast on and fast off kinetics to help capture cells out of the circulating shear flow; subsequently, tight binding by dimeric/oligomeric selectins is enabled to significantly slow rolling. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.en
dc.description.sponsorshipKAUST, King Abdullah University of Science and Technologyen
dc.publisherAmerican Society for Biochemistry & Molecular Biology (ASBMB)en
dc.titleQuantitative Characterization of E-selectin Interaction with Native CD44 and P-selectin Glycoprotein Ligand-1 (PSGL-1) Using a Real Time Immunoprecipitation-based Binding Assayen
dc.typeArticleen
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Divisionen
dc.identifier.journalJournal of Biological Chemistryen
dc.identifier.pmcidPMC4571854en
kaust.authorAbu Samra, Dina Bashir Kamilen
kaust.authorAl Kilani, Aliaen
kaust.authorHamdan, Samiren
kaust.authorSakashita, Kosukeen
kaust.authorGadhoum, Samahen
kaust.authorMerzaban, Jasmeen S.en

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