6URP

RDC refined solution structure of the insecticidal toxin Ta1a

6URP の概要

• エントリーDOI: 10.2210/pdb6urp/pdb
• NMR情報: BMRB: 16667
• 分子名称: U1-agatoxin-Ta1a (1 entity in total)
• 機能のキーワード: disulfide-rich peptides, peptides, residual dipolar couplings, rdcs, toxin
• 由来する生物種: Eratigena agrestis (Hobo spider)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 5781.39

構造登録者

Ramanujam, V.,Shen, Y.,Ying, J.,Mobli, M. (登録日: 2019-10-24, 公開日: 2020-01-22, 最終更新日: 2024-10-23)

主引用文献

Ramanujam, V.,Shen, Y.,Ying, J.,Mobli, M.
Residual Dipolar Couplings for Resolving Cysteine Bridges in Disulfide-Rich Peptides.
Front Chem, 7:889-889, 2019

PubMed Abstract: Disulfide bridges in proteins are formed by the oxidation of pairs of cysteine residues. These cross-links play a critical role in stabilizing the 3D-structure of small disulfide rich polypeptides such as hormones and venom toxins. The arrangement of the multiple disulfide bonds directs the peptide fold into distinct structural motifs that have evolved for resistance against biochemical and physical insults. These structural scaffolds have, therefore, proven to be very attractive in bioengineering efforts to develop novel biologics with applications in health and agriculture. Structural characterization of small disulfide rich peptides (DRPs) presents unique challenges when using commonly applied biophysical methods. NMR is the most commonly used method for studying such molecules, where the relatively small size of these molecules results in highly precise structural ensembles defined by a large number of distance and dihedral angle restraints per amino acid. However, in NMR the sulfur atoms that are involved in three of the five dihedral angles in a disulfide bond cannot be readily measured. Given the central role of disulfide bonds in the structure of these molecules, it is unclear what the inherent resolution of such NMR structures is when using traditional NMR methods. Here, we use an extensive set of long-range residual dipolar couplings (RDCs) to assess the resolution of the NMR structure of a disulfide-rich peptide. We find that structures based primarily on NOEs, yield ensembles that are equivalent to a crystallographic resolution of 2-3 Å in resolution, and that incorporation of RDCs reduces this to ~1-1.5 Å resolution. At this resolution the sidechain of ordered amino acids can be defined accurately, allowing the geometry of the cysteine bridges to be better defined, and allowing for disulfide-bond connectivities to be determined with high confidence. The observed improvements in resolution when using RDCs is remarkable considering the small size of these peptides.

PubMed: 32039137
DOI: 10.3389/fchem.2019.00889

実験手法

SOLUTION NMR