The solution structure of EMILIN1 globular C1q domain reveals a disordered insertion necessary for interaction with the α4β1 integrin

Giuliana Verdone, Roberto Doliana, Alessandra Corazza, Simon A. Colebrooke, Paola Spessotto, Simonetta Bot, Francesco Bucciotti, Alessandra Capuano, Alessandra Silvestri, Paolo Viglino, Iain D. Campbell, Alfonso Colombatti, Gennaro Esposito

Research output: Contribution to journalArticle

Abstract

The extracellular matrix protein EMILIN1 (elastin microfibril interface located protein 1) is implicated in maintaining blood pressure homeostasis via the N-terminal elastin microfibril interface domain and in trophoblast invasion of the uterine wall via the globular C1q (gC1q) domain. Here, we describe the first NMR-based homology model structure of the human 52-kDa homotrimer of the EMILIN1 gC1q domain. In contrast to all of the gC1q (crystal) structures solved to date, the 10-stranded β-sandwich fold of the gC1q domain is reduced to nine β strands with a consequent increase in the size of the central cavity lumen. An unstructured loop, resulting from an insertion unique to EMILIN1 and EMILIN2 family members and located at the trimer apex upstream of the missing strand, specifically engages the α4β1 integrin. Using both Jurkat T and EA.hy926 endothelial cells as well as site-directed mutagenesis, we demonstrate that the ability of α4β1 integrins to recognize the trimeric EMILIN1 gC1q domain mainly depends on a single glutamic acid residue (Glu933). Static and flow adhesion of T cells and haptotactic migration of endothelial cells on gC1q is fully dependent on this residue. Thus, EMILIN1 gC1q-α4β1 represents a unique ligand/receptor system, with a requirement for a 3-fold arrangement of the interaction site.

Original languageEnglish (US)
Pages (from-to)18947-18956
Number of pages10
JournalJournal of Biological Chemistry
Volume283
Issue number27
DOIs
StatePublished - Jul 4 2008

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Integrins
Endothelial cells
Endothelial Cells
Microfibrils
Mutagenesis
Aptitude
T-cells
Elastin
Extracellular Matrix Proteins
Blood pressure
Trophoblasts
Model structures
Site-Directed Mutagenesis
Cell Movement
Glutamic Acid
Homeostasis
Adhesion
Crystal structure
Nuclear magnetic resonance
elastin microfibril interface located protein

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

The solution structure of EMILIN1 globular C1q domain reveals a disordered insertion necessary for interaction with the α4β1 integrin. / Verdone, Giuliana; Doliana, Roberto; Corazza, Alessandra; Colebrooke, Simon A.; Spessotto, Paola; Bot, Simonetta; Bucciotti, Francesco; Capuano, Alessandra; Silvestri, Alessandra; Viglino, Paolo; Campbell, Iain D.; Colombatti, Alfonso; Esposito, Gennaro.

In: Journal of Biological Chemistry, Vol. 283, No. 27, 04.07.2008, p. 18947-18956.

Research output: Contribution to journalArticle

Verdone, G, Doliana, R, Corazza, A, Colebrooke, SA, Spessotto, P, Bot, S, Bucciotti, F, Capuano, A, Silvestri, A, Viglino, P, Campbell, ID, Colombatti, A & Esposito, G 2008, 'The solution structure of EMILIN1 globular C1q domain reveals a disordered insertion necessary for interaction with the α4β1 integrin', Journal of Biological Chemistry, vol. 283, no. 27, pp. 18947-18956. https://doi.org/10.1074/jbc.M801085200
Verdone, Giuliana ; Doliana, Roberto ; Corazza, Alessandra ; Colebrooke, Simon A. ; Spessotto, Paola ; Bot, Simonetta ; Bucciotti, Francesco ; Capuano, Alessandra ; Silvestri, Alessandra ; Viglino, Paolo ; Campbell, Iain D. ; Colombatti, Alfonso ; Esposito, Gennaro. / The solution structure of EMILIN1 globular C1q domain reveals a disordered insertion necessary for interaction with the α4β1 integrin. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 27. pp. 18947-18956.
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AU - Colebrooke, Simon A.

AU - Spessotto, Paola

AU - Bot, Simonetta

AU - Bucciotti, Francesco

AU - Capuano, Alessandra

AU - Silvestri, Alessandra

AU - Viglino, Paolo

AU - Campbell, Iain D.

AU - Colombatti, Alfonso

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N2 - The extracellular matrix protein EMILIN1 (elastin microfibril interface located protein 1) is implicated in maintaining blood pressure homeostasis via the N-terminal elastin microfibril interface domain and in trophoblast invasion of the uterine wall via the globular C1q (gC1q) domain. Here, we describe the first NMR-based homology model structure of the human 52-kDa homotrimer of the EMILIN1 gC1q domain. In contrast to all of the gC1q (crystal) structures solved to date, the 10-stranded β-sandwich fold of the gC1q domain is reduced to nine β strands with a consequent increase in the size of the central cavity lumen. An unstructured loop, resulting from an insertion unique to EMILIN1 and EMILIN2 family members and located at the trimer apex upstream of the missing strand, specifically engages the α4β1 integrin. Using both Jurkat T and EA.hy926 endothelial cells as well as site-directed mutagenesis, we demonstrate that the ability of α4β1 integrins to recognize the trimeric EMILIN1 gC1q domain mainly depends on a single glutamic acid residue (Glu933). Static and flow adhesion of T cells and haptotactic migration of endothelial cells on gC1q is fully dependent on this residue. Thus, EMILIN1 gC1q-α4β1 represents a unique ligand/receptor system, with a requirement for a 3-fold arrangement of the interaction site.

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