Structure of the putative 32 kDa myrosinase-binding protein from Arabidopsis (At3g16450.1) determined by SAIL-NMR.
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.
Takeda
Mitsuhiro
M
Graduate School of Science, Nagoya University, Japan.
Sugimori
Nozomi
N
Torizawa
Takuya
T
Terauchi
Tsutomu
T
Ono
Akira M
AM
Yagi
Hirokazu
H
Yamaguchi
Yoshiki
Y
Kato
Koichi
K
Ikeya
Teppei
T
Jee
Jungoo
J
Güntert
Peter
P
Aceti
David J
DJ
Markley
John L
JL
Kainosho
Masatsune
M
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
D029681:Arabidopsis Proteins; D006023:Glycoproteins; D047908:Intracellular Signaling Peptides and Proteins; C099049:MyAP protein, Brassica napus; D009844:Oligosaccharides; D010940:Plant Proteins; D011994:Recombinant Proteins