Premium
Structure of the putative 32 kDa myrosinase‐binding protein from Arabidopsis (At3g16450.1) determined by SAIL‐NMR
Author(s) -
Takeda Mitsuhiro,
Sugimori Nozomi,
Torizawa Takuya,
Terauchi Tsutomu,
Ono Akira M.,
Yagi Hirokazu,
Yamaguchi Yoshiki,
Kato Koichi,
Ikeya Teppei,
Jee JunGoo,
Güntert Peter,
Aceti David J.,
Markley John L.,
Kainosho Masatsune
Publication year - 2008
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2008.06717.x
Subject(s) - myrosinase , arabidopsis , glucosinolate , arabidopsis thaliana , biochemistry , heteronuclear single quantum coherence spectroscopy , biology , chemistry , two dimensional nuclear magnetic resonance spectroscopy , stereochemistry , gene , botany , mutant , brassica
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 m m ) 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 β‐sheets.