title: Zebavidin--an avidin-like protein from zebrafish.
abstract: The avidin protein family members are well known for their high affinity towards D-biotin and high structural stability. These properties make avidins valuable tools for a wide range of biotechnology applications. We have identified a new member of the avidin family in the zebrafish (Danio rerio) genome, hereafter called zebavidin. The protein is highly expressed in the gonads of both male and female zebrafish and in the gills of male fish, but our data suggest that zebavidin is not crucial for the developing embryo. Biophysical and structural characterisation of zebavidin revealed distinct properties not found in any previously characterised avidins. Gel filtration chromatography and native mass spectrometry suggest that the protein forms dimers in the absence of biotin at low ionic strength, but assembles into tetramers upon binding biotin. Ligand binding was analysed using radioactive and fluorescently labelled biotin and isothermal titration calorimetry. Moreover, the crystal structure of zebavidin in complex with biotin was solved at 2.4 Å resolution and unveiled unique ligand binding and subunit interface architectures; the atomic-level details support our physicochemical observations.
authors.affiliation: Institute of Biomedical Technology, University of Tampere, BioMediTech, Tampere, Finland ; Fimlab Laboratories, Pirkanmaa Hospital District, Tampere, Finland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Taskinen
authors.forename: Joanna
authors.initials: J
authors.lastname: Zmurko
authors.forename: Markus
authors.initials: M
authors.lastname: Ojanen
authors.forename: Sampo
authors.initials: S
authors.lastname: Kukkurainen
authors.forename: Marimuthu
authors.initials: M
authors.lastname: Parthiban
authors.forename: Juha A E
authors.initials: JA
authors.lastname: Määttä
authors.forename: Jenni
authors.initials: J
authors.lastname: Leppiniemi
authors.forename: Janne
authors.initials: J
authors.lastname: Jänis
authors.forename: Mataleena
authors.initials: M
authors.lastname: Parikka
authors.forename: Hannu
authors.initials: H
authors.lastname: Turpeinen
authors.forename: Mika
authors.initials: M
authors.lastname: Rämet
authors.forename: Marko
authors.initials: M
authors.lastname: Pesu
authors.forename: Mark S
authors.initials: MS
authors.lastname: Johnson
authors.forename: Markku S
authors.initials: MS
authors.lastname: Kulomaa
authors.forename: Tomi T
authors.initials: TT
authors.lastname: Airenne
authors.forename: Vesa P
authors.initials: VP
authors.lastname: Hytönen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001360:Avidin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D004625:Embryo, Nonmammalian; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D029941:Fish Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D016678:Genome; D005880:Gills / Q000196:embryology / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006066:Gonads / Q000196:embryology / Q000378:metabolism; D008297:Male; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism; D029961:Zebrafish Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D029941:Fish Proteins; D006023:Glycoproteins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D029961:Zebrafish Proteins; C000590111:avd protein, zebrafish; D001360:Avidin; D001710:Biotin