title: The highly dynamic oligomeric structure of bradavidin II is unique among avidin proteins.
abstract: Bradavidin II is a biotin-binding protein from Bradyrhizobium japonicum that resembles chicken avidin and bacterial streptavidin. A biophysical characterization was carried out using dynamic light scattering, native mass spectrometry, differential scanning calorimetry, and isothermal titration calorimetry combined with structural characterization using X-ray crystallography. These observations revealed that bradavidin II differs from canonical homotetrameric avidin protein family members in its quaternary structure. In contrast with the other avidins, bradavidin II appears to have a dynamic (transient) oligomeric state in solution. It is monomeric at low protein concentrations but forms higher oligomeric assemblies at higher concentrations. The crystal structure of bradavidin II revealed an important role for Phe42 in shielding the bound ligand from surrounding water molecules, thus functionally replacing the L7,8 loop essential for tight ligand binding in avidin and streptavidin. This bradavidin II characterization opens new avenues for oligomerization-independent biotin-binding protein development.
authors: Institute of Biomedical Technology, University of Tampere and Tampere University Hospital, FI-33014, Tampere, Finland.
authors: Jenni
authors: J
authors: Leppiniemi
authors: Amit
authors: A
authors: Meir
authors: Niklas
authors: N
authors: Kähkönen
authors: Sampo
authors: S
authors: Kukkurainen
authors: Juha A
authors: JA
authors: Määttä
authors: Markus
authors: M
authors: Ojanen
authors: Janne
authors: J
authors: Jänis
authors: Markku S
authors: MS
authors: Kulomaa
authors: Oded
authors: O
authors: Livnah
authors: Vesa P
authors: VP
authors: Hytönen
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry* / Q000378:metabolism*; D002645:Chickens; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D055503:Protein Multimerization; D058767:Protein Unfolding; D016415:Sequence Alignment; D013696:Temperature
chemical_list: D002352:Carrier Proteins; C501304:bradavidin protein, Bradyrhizobium japonicum; D001710:Biotin
references: Biophys J. 2004 Oct;87(4):2701-13
references: 15298874
references: Acta Crystallogr D Biol Crystallogr. 2005 May;61(Pt 5):528-38
references: 15858262
references: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2184-95
references: 15572771
references: J Mol Biol. 2009 Feb 20;386(2):379-90
references: 19111749
references: Methods Mol Biol. 2008;426:419-35
references: 18542881
references: PLoS One. 2012;7(5):e35962
references: 22574129
references: Biochim Biophys Acta. 2008 Jul-Aug;1784(7-8):1002-10
references: 18486632
references: J Mol Biol. 1998 May 29;279(1):211-21
references: 9636711
references: Biochem J. 2007 Aug 1;405(3):397-405
references: 17447892
references: J Appl Crystallogr. 2007 Aug 1;40(Pt 4):658-674
references: 19461840
references: Protein Sci. 2000 May;9(5):878-85
references: 10850797
references: Proc Natl Acad Sci U S A. 1995 Apr 11;92(8):3180-4
references: 7724536
references: Cell Mol Life Sci. 2006 Dec;63(24):2992-3017
references: 17086379
references: J Biol Chem. 2012 May 25;287(22):17951-62
references: 22493427
references: Methods Enzymol. 2003;374:229-44
references: 14696376
references: BMC Struct Biol. 2009 Sep 29;9:63
references: 19788720
references: Protein Sci. 2012 Feb;21(2):211-8
references: 22109962
references: Biotechnol Bioeng. 2013 Jan;110(1):57-67
references: 22806584
references: BMC Biotechnol. 2005 Oct 07;5:28
references: 16212654
references: Methods Enzymol. 1990;184:51-67
references: 2388586
references: J Biol Chem. 2003 Feb 7;278(6):4010-4
references: 12458212
references: Biochemistry. 2011 Oct 11;50(40):8682-91
references: 21892837
references: J Biol Chem. 2005 Apr 8;280(14):13250-5
references: 15695809
references: Methods Enzymol. 1997;276:307-26
references: 27754618
references: J Biol Chem. 2004 Mar 5;279(10):9337-43
references: 14660583
references: Biochem Biophys Res Commun. 2004 Dec 17;325(3):922-7
references: 15541378
references: Nucleic Acids Res. 2009 Jan;37(Database issue):D387-92
references: 18931379
references: Chem Biol. 2006 Oct;13(10):1029-39
references: 17052607
references: Anal Chem. 2013 Feb 5;85(3):1591-6
references: 23252501
references: FEBS J. 2009 Mar;276(5):1383-97
references: 19187241
references: Biochemistry. 1995 Sep 19;34(37):11750-6
references: 7547907
references: Electrophoresis. 1996 Aug;17(8):1319-24
references: 8874057
references: Biochem J. 2003 Jan 15;369(Pt 2):249-54
references: 12358604
references: Proc Natl Acad Sci U S A. 1993 Jun 1;90(11):5076-80
references: 8506353
references: Biochem J. 2004 Dec 1;384(Pt 2):385-90
references: 15324300
references: Bioinformatics. 2006 Jan 15;22(2):195-201
references: 16301204
references: Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
references: 15572765
references: Biotechnol Bioeng. 2011 Mar;108(3):481-90
references: 20939005
references: Anal Biochem. 2000 Jan 15;277(2):260-6
references: 10625516
references: Adv Protein Chem. 1975;29:85-133
references: 237414
references: Trends Biotechnol. 2007 Jun;25(6):269-77
references: 17433846