title: Structure of the putative 32 kDa myrosinase-binding protein from Arabidopsis (At3g16450.1) determined by SAIL-NMR.
abstract: The product of gene At3g16450.1 from Arabidopsis thaliana is a 32 kDa, 299-residue protein classified as resembling a myrosinase-binding protein (MyroBP). MyroBPs are found in plants as part of a complex with the glucosinolate-degrading enzyme myrosinase, and are suspected to play a role in myrosinase-dependent defense against pathogens. Many MyroBPs and MyroBP-related proteins are composed of repeated homologous sequences with unknown structure. We report here the three-dimensional structure of the At3g16450.1 protein from Arabidopsis, which consists of two tandem repeats. Because the size of the protein is larger than that amenable to high-throughput analysis by uniform (13)C/(15)N labeling methods, we used stereo-array isotope labeling (SAIL) technology to prepare an optimally (2)H/(13)C/(15)N-labeled sample. NMR data sets collected using the SAIL protein enabled us to assign (1)H, (13)C and (15)N chemical shifts to 95.5% of all atoms, even at a low concentration (0.2 mm) of protein product. We collected additional NOESY data and determined the three-dimensional structure using the cyana software package. The structure, the first for a MyroBP family member, revealed that the At3g16450.1 protein consists of two independent but similar lectin-fold domains, each composed of three beta-sheets.
authors.affiliation: Graduate School of Science, Nagoya University, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Takeda
authors.forename: Nozomi
authors.initials: N
authors.lastname: Sugimori
authors.forename: Takuya
authors.initials: T
authors.lastname: Torizawa
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Terauchi
authors.forename: Akira M
authors.initials: AM
authors.lastname: Ono
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Yagi
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Yamaguchi
authors.forename: Koichi
authors.initials: K
authors.lastname: Kato
authors.forename: Teppei
authors.initials: T
authors.lastname: Ikeya
authors.forename: Jungoo
authors.initials: J
authors.lastname: Jee
authors.forename: Peter
authors.initials: P
authors.lastname: Güntert
authors.forename: David J
authors.initials: DJ
authors.lastname: Aceti
authors.forename: John L
authors.initials: JL
authors.lastname: Markley
authors.forename: Masatsune
authors.initials: M
authors.lastname: Kainosho
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins; D007553:Isotope Labeling / Q000379:methods; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry
chemical_list: D029681:Arabidopsis Proteins; D006023:Glycoproteins; D047908:Intracellular Signaling Peptides and Proteins; C099049:MyAP protein, Brassica napus; D009844:Oligosaccharides; D010940:Plant Proteins; D011994:Recombinant Proteins