1.8 Å structure of murine GITR ligand dimer expressed in Drosophila melanogaster S2 cells

Kausik Chattopadhyay, Udupi A. Ramagopal, Stanley G. Nathenson, Steven C. Almo

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Glucocorticoid-induced TNF receptor ligand (GITRL), a prominent member of the TNF superfamily, activates its receptor on both effector and regulatory T cells to generate critical costimulatory signals that have been implicated in a wide range of T-cell immune functions. The crystal structures of murine and human orthologs of GITRL recombinantly expressed in Escherichia coli have previously been determined. In contrast to all classical TNF structures, including the human GITRL structure, murine GITRL demonstrated a unique 'strand-exchanged' dimeric organization. Such a novel assembly behavior indicated a dramatic impact on receptor activation as well as on the signaling mechanism associated with the murine GITRL costimulatory system. In this present work, the 1.8 Å resolution crystal structure of murine GITRL expressed in Drosophila melanogaster S2 cells is reported. The eukaryotic protein-expression system allows transport of the recombinant protein into the extracellular culture medium, thus maximizing the possibility of obtaining correctly folded material devoid of any folding/assembly artifacts that are often suspected with E. coli-expressed proteins. The S2 cell-expressed murine GITRL adopts an identical 'strand-exchanged' dimeric structure to that observed for the E. coli-expressed protein, thus conclusively demonstrating the novel quaternary structure assembly behavior of murine GITRL.

Original languageEnglish (US)
Pages (from-to)434-439
Number of pages6
JournalActa Crystallographica Section D: Biological Crystallography
Volume65
Issue number5
DOIs
StatePublished - Apr 18 2009

Fingerprint

Tumor Necrosis Factors
Drosophila melanogaster
Glucocorticoids
Ligands
Escherichia coli Proteins
Regulatory T-Lymphocytes
Recombinant Proteins
Artifacts
Culture Media
Carrier Proteins
Escherichia coli
T-Lymphocytes

Keywords

  • Expression in Drosophila melanogaster
  • Glucocorticoid-induced TNF receptor ligand

ASJC Scopus subject areas

  • Structural Biology
  • Medicine(all)

Cite this

1.8 Å structure of murine GITR ligand dimer expressed in Drosophila melanogaster S2 cells. / Chattopadhyay, Kausik; Ramagopal, Udupi A.; Nathenson, Stanley G.; Almo, Steven C.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 65, No. 5, 18.04.2009, p. 434-439.

Research output: Contribution to journalArticle

Chattopadhyay, Kausik ; Ramagopal, Udupi A. ; Nathenson, Stanley G. ; Almo, Steven C. / 1.8 Å structure of murine GITR ligand dimer expressed in Drosophila melanogaster S2 cells. In: Acta Crystallographica Section D: Biological Crystallography. 2009 ; Vol. 65, No. 5. pp. 434-439.
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