title: Creation of a S1P Lyase bacterial surrogate for structure-based drug design.
abstract: S1P Lyase (SPL) has been described as a drug target in the treatment of autoimmune diseases. It plays an important role in maintaining intracellular levels of S1P thereby affecting T cell egress from lymphoid tissues. Several groups have already published approaches to inhibit S1P Lyase with small molecules, which in turn increase endogenous S1P concentrations resulting in immunosuppression. The use of structural biology has previously aided SPL inhibitor design. Novel construct design is at times necessary to provide a reagent for protein crystallography. Here we present a chimeric bacterial protein scaffold used for protein X-ray structures in the presence of early small molecule inhibitors. Mutations were introduced to the bacterial SPL from Symbiobacterium thermophilum which mimic the human enzyme. As a result, two mutant StSPL crystal structures resolved to 2.8Å and 2.2Å resolutions were solved and provide initial structural hypotheses for an isoxazole chemical series, whose optimization is discussed in the accompanying paper.
authors.affiliation: AbbVie Bioresearch Center, 381 Plantation Street, Worcester, MA 01605, USA. Electronic address: maria.argiriadi@abbvie.com.
authors.forename: Maria A
authors.initials: MA
authors.lastname: Argiriadi
authors.affiliation: AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
authors.forename: David
authors.initials: D
authors.lastname: Banach
authors.affiliation: AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
authors.forename: Elzbieta
authors.initials: E
authors.lastname: Radziejewska
authors.affiliation: AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
authors.forename: Susan
authors.initials: S
authors.lastname: Marchie
authors.affiliation: AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: DiMauro
authors.affiliation: AbbVie, 1 North Waukegan Road, North Chicago, IL 60064, USA.
authors.forename: Jurgen
authors.initials: J
authors.lastname: Dinges
authors.affiliation: AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Dominguez
authors.affiliation: AbbVie, 1 North Waukegan Road, North Chicago, IL 60064, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Hutchins
authors.affiliation: AbbVie, 1 North Waukegan Road, North Chicago, IL 60064, USA.
authors.forename: Russell A
authors.initials: RA
authors.lastname: Judge
authors.affiliation: AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
authors.forename: Kara
authors.initials: K
authors.lastname: Queeney
authors.affiliation: AbbVie Bioresearch Center, 381 Plantation Street, Worcester, MA 01605, USA.
authors.forename: Grier
authors.initials: G
authors.lastname: Wallace
authors.affiliation: AbbVie Bioresearch Center, 100 Research Drive, Worcester, MA 01605, USA.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Harris
mesh_terms: D000446:Aldehyde-Lyases / Q000737:chemistry / Q000378:metabolism*; D018360:Crystallography, X-Ray; D015195:Drug Design; D004926:Escherichia coli / Q000201:enzymology*
chemical_list: D000446:Aldehyde-Lyases; C022649:sphingosine 1-phosphate lyase (aldolase)
keywords: Crystal structure; Homology modeling; S1P Lyase; Site-directed mutagenesis