title: High level expression of the Drosophila Toll receptor ectodomain and crystallization of its complex with the morphogen Spätzle.
abstract: Drosophila Toll receptors are involved in embryonic development and in the immune response of adult flies. In both processes, the Toll receptor ligand is the NGF-like cystine knot protein Spätzle. Here we present the expression of Toll receptor ectodomain in Schneider cells at high yields and demonstrate a high affinity interaction with the refolded and trypsin-processed Spätzle cystine knot domain dimer. Poorly and anisotropically diffracting crystals of the complex could be improved by deglycosylation and dehydration, paving the way for structural analyses of the Toll-Spätzle interaction.
authors.affiliation: Martin-Luther- Universität Halle-Wittenberg, Institut f u r Biotechnologie, Kurt-Mothes-Strasse 3, D-06120 Halle Saale, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Stelter
authors.forename: Uwe
authors.initials: U
authors.lastname: Fandrich
authors.forename: Kati
authors.initials: K
authors.lastname: Franzke
authors.forename: Angelika
authors.initials: A
authors.lastname: Schierhorn
authors.forename: Constanze
authors.initials: C
authors.lastname: Breithaupt
authors.forename: Christoph
authors.initials: C
authors.lastname: Parthier
authors.forename: Milton T
authors.initials: MT
authors.lastname: Stubbs
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D003460:Crystallization; D004330:Drosophila / Q000737:chemistry / Q000378:metabolism*; D029721:Drosophila Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D008969:Molecular Sequence Data; D055503:Protein Multimerization; D058849:Protein Refolding; D017434:Protein Structure, Tertiary; D051193:Toll-Like Receptors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D029721:Drosophila Proteins; C068721:Tl protein, Drosophila; D051193:Toll-Like Receptors; C086179:spz protein, Drosophila