title: Identification of Avian Corticosteroid-binding Globulin (SerpinA6) Reveals the Molecular Basis of Evolutionary Adaptations in SerpinA6 Structure and Function as a Steroid-binding Protein.
abstract: Corticosteroid-binding globulin (CBG) was isolated from chicken serum and identified by mass spectrometry and genomic analysis. This revealed that the organization and synteny of avian and mammalian SerpinA6 genes are conserved. Recombinant zebra finch CBG steroid-binding properties reflect those of the natural protein in plasma and confirm its identity. Zebra finch and rat CBG crystal structures in complex with cortisol resemble each other, but their primary structures share only ∼40% identity, and their steroid-binding site topographies differ in several unexpected ways. Remarkably, a tryptophan that anchors ligands in mammalian CBG steroid-binding sites is replaced by an asparagine. Phylogenetic comparisons show that reptilian CBG orthologs share this unexpected property. Glycosylation of this asparagine in zebra finch CBG does not influence its steroid-binding affinity, but we present evidence that it may participate in protein folding and steroid-binding site formation. Substitutions of amino acids within zebra finch CBG that are conserved only in birds reveal how they contribute to their distinct steroid-binding properties, including their high (nanomolar) affinities for glucocorticoids, progesterone, and androgens. As in mammals, a protease secreted by Pseudomonas aeruginosa cleaves CBG in zebra finch plasma within its reactive center loop and disrupts steroid binding, suggesting an evolutionarily conserved property of CBGs. Measurements of CBG mRNA in zebra finch tissues indicate that liver is the main site of plasma CBG production, and anti-zebra finch CBG antibodies cross-react with CBGs in other birds, extending opportunities to study how CBG regulates the actions of glucocorticoids and sex steroids in these species.
authors.affiliation: From the Departments of Cellular and Physiological Sciences.
authors.forename: Ganna
authors.initials: G
authors.lastname: Vashchenko
authors.affiliation: Biochemistry and Molecular Biology, and.
authors.forename: Samir
authors.initials: S
authors.lastname: Das
authors.affiliation: Biochemistry and Molecular Biology, and.
authors.forename: Kyung-Mee
authors.initials: KM
authors.lastname: Moon
authors.affiliation: Biochemistry and Molecular Biology, and.
authors.forename: Jason C
authors.initials: JC
authors.lastname: Rogalski
authors.affiliation: Psychology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Taves
authors.affiliation: Psychology, University of British Columbia, Vancouver, British Columbia V6T 1Z3, Canada.
authors.forename: Kiran K
authors.initials: KK
authors.lastname: Soma
authors.affiliation: Biochemistry and Molecular Biology, and.
authors.forename: Filip
authors.initials: F
authors.lastname: Van Petegem
authors.affiliation: Biochemistry and Molecular Biology, and.
authors.forename: Leonard J
authors.initials: LJ
authors.lastname: Foster
authors.affiliation: From the Departments of Cellular and Physiological Sciences, geoffrey.hammond@ubc.ca.
authors.forename: Geoffrey L
authors.initials: GL
authors.lastname: Hammond
mesh_terms: D000222:Adaptation, Physiological; D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins / Q000097:blood* / Q000737:chemistry / Q000235:genetics*; D001717:Birds / Q000097:blood* / Q000235:genetics*; D002645:Chickens / Q000097:blood / Q000235:genetics; D018360:Crystallography, X-Ray; D019143:Evolution, Molecular; D046369:Finches / Q000097:blood / Q000235:genetics; D006031:Glycosylation; D008958:Models, Molecular; D010802:Phylogeny; D051381:Rats; D017386:Sequence Homology, Amino Acid; D046370:Sparrows / Q000097:blood / Q000235:genetics; D014156:Transcortin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
chemical_list: D030161:Avian Proteins; D014156:Transcortin
keywords: N-linked glycosylation; androgen; birds; glucocorticoid; progesterone; protein crystallization; serine proteinase inhibitor; stress