Functional Binding of E-selectin to its Ligands is Enhanced by Structural Features Beyond its Lectin Domain
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ArticleAuthors
Aleisa, Fajr A
Sakashita, Kosuke

Lee, Jae Man
Abu Samra, Dina Bashir Kamil

Al Alwan, Bader

Nozue, Shuho
Tehseen, Muhammad

Hamdan, Samir

Habuchi, Satoshi

Kusakabe, Takahiro
Merzaban, Jasmeen

KAUST Department
Biological and Environmental Sciences and Engineering (BESE) DivisionBioscience Program
Chemical Engineering Program
Laboratory of DNA Replication and Recombination
Physical Science and Engineering (PSE) Division
Proteomics, protein expression & cytomet
Single-Molecule Spectroscopy and Microscopy Research Group
KAUST Grant Number
OCRF-2014-CRG3-2276Date
2020-01-16Online Publication Date
2020-01-16Print Publication Date
2020-03-13Submitted Date
2019-09-19Permanent link to this record
http://hdl.handle.net/10754/661071
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Show full item recordAbstract
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.Citation
Aleisa, F. A., Sakashita, K., Lee, J. M., AbuSamra, D. B., Alwan, B., Nozue, S., … Merzaban, J. S. (2020). Functional Binding of E-selectin to its Ligands is Enhanced by Structural Features Beyond its Lectin Domain. Journal of Biological Chemistry, jbc.RA119.010910. doi:10.1074/jbc.ra119.010910Sponsors
This research was supported by a King Abdullah University of Science and Technology (KAUST) Faculty Baseline Research Funding Program to J.S.M. and by a Competitive Research Grant (OCRF-2014-CRG3-2276) awarded to J.S.M. We would like to express our gratitude to Mr. Vlad-Stefan Raducanu for discussion about E-S0 sample aggregates; Ms. Maryam Mih, Ms. Samar A. Rustum and Ms. Umme Habiba for their support in the management of the lab. We would also like to thank Veronica Tremblay from the KAUST Academic Writing Service for editing the manuscript.Journal
Journal of Biological ChemistryAdditional Links
http://www.jbc.org/lookup/doi/10.1074/jbc.RA119.010910ae974a485f413a2113503eed53cd6c53
10.1074/jbc.ra119.010910