Functional Binding of E-selectin to its Ligands is Enhanced by Structural Features Beyond its Lectin Domain

Fajr A Aleisa, Kosuke Sakashita, Jae Man Lee, Dina Bashir Kamil Abu Samra, Bader Al Alwan, Shuho Nozue, Muhammad Tehseen, Samir Hamdan, Satoshi Habuchi, Takahiro Kusakabe, Jasmeen Merzaban

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

Selectins are key to mediating interactions involved in cellular adhesion and migration, underlying processes such as immune responses, metastasis, and transplantation. Selectins are composed of a lectin domain, an epidermal growth factor (EGF)-like domain, multiple short consensus repeats (SCRs), a transmembrane domain, and a cytoplasmic tail. It is well established that the lectin and EGF domains are required to mediate interactions with ligands; however, the contributions of the other domains in mediating these interactions remain obscure. Using various E-selectin constructs produced in a newly developed silkworm-based expression system and several assays performed under both static and physiological flow conditions, including flow cytometry, glycan array analysis, surface plasmon resonance, and cell-rolling assays, we show here that a reduction in the number of SCR domains is correlated with a decline in functional E-selectin binding to hematopoietic cell E- or L- selectin ligand (HCELL) and P-selectin glycoprotein ligand-1 (PSGL-1). Moreover, the binding was significantly improved through E-selectin dimerization and by a substitution (A28H) that mimics an extended conformation of the lectin and EGF domains. Analyses of the association and dissociation rates indicated that the SCR domains, conformational extension, and dimerization collectively contribute to the association rate of E-selectin–ligand binding, whereas just the lectin and EGF domains contribute to the dissociation rate. These findings provide the first evidence of the critical role of the association rate in functional E-selectin–ligand interactions, and they highlight that the SCR domains have an important role that goes beyond the structural extension of the lectin and EGF domains.
Original languageEnglish (US)
Pages (from-to)jbc.RA119.010910
JournalJournal of Biological Chemistry
Volume295
Issue number11
DOIs
StatePublished - Jan 16 2020

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