8ULM

Chickpea (Cicer arientinum) nodule-specific cysteine-rich peptide NCR13: Solution NMR structure of the isomer with C4:C23, C15:C30, and C10:C28 disulfide bonds

8ULM の概要

• エントリーDOI: 10.2210/pdb8ulm/pdb
• 関連するPDBエントリー: 7th8
• NMR情報: BMRB: 31111
• 分子名称: Nodule cysteine-rich protein 13 (1 entity in total)
• 機能のキーワード: antifunal activity, endosymbiotic, disulfide-rich, antimicrobial protein, ncr peptide, antifungal protein
• 由来する生物種: Cicer arietinum (chickpea)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 3741.61

構造登録者

Buchko, G.W.,Zhou, M.,Shah, D.M.,Velivelli, S.L.S. (登録日: 2023-10-16, 公開日: 2023-11-01, 最終更新日: 2024-12-25)

主引用文献

Godwin, J.,Djami-Tchatchou, A.T.,Velivelli, S.L.S.,Tetorya, M.,Kalunke, R.,Pokhrel, A.,Zhou, M.,Buchko, G.W.,Czymmek, K.J.,Shah, D.M.
Chickpea NCR13 disulfide cross-linking variants exhibit profound differences in antifungal activity and modes of action.
Plos Pathog., 20:e1012745-e1012745, 2024

PubMed Abstract: Small cysteine-rich antifungal peptides with multi-site modes of action (MoA) have potential for development as biofungicides. In particular, legumes of the inverted repeat-lacking clade express a large family of nodule-specific cysteine-rich (NCR) peptides that orchestrate differentiation of nitrogen-fixing bacteria into bacteroids. These NCRs can form two or three intramolecular disulfide bonds and a subset of these peptides with high cationicity exhibits antifungal activity. However, the importance of intramolecular disulfide pairing and MoA against fungal pathogens for most of these plant peptides remains to be elucidated. Our study focused on a highly cationic chickpea NCR13, which has a net charge of +8 and contains six cysteines capable of forming three disulfide bonds. NCR13 expression in Pichia pastoris resulted in formation of two peptide folding variants, NCR13_PFV1 and NCR13_PFV2, that differed in the pairing of two out of three disulfide bonds despite having an identical amino acid sequence. The NMR structure of each PFV revealed a unique three-dimensional fold with the PFV1 structure being more compact but less dynamic. Surprisingly, PFV1 and PFV2 differed profoundly in the potency of antifungal activity against several fungal plant pathogens and their multi-faceted MoA. PFV1 showed significantly faster fungal cell-permeabilizing and cell entry capabilities as well as greater stability once inside the fungal cells. Additionally, PFV1 was more effective in binding fungal ribosomal RNA and inhibiting protein translation in vitro. Furthermore, when sprayed on pepper and tomato plants, PFV1 was more effective in reducing disease symptoms caused by Botrytis cinerea, causal agent of gray mold disease in fruits, vegetables, and flowers. In conclusion, our work highlights the significant impact of disulfide pairing on the antifungal activity and MoA of NCR13 and provides a structural framework for design of novel, potent antifungal peptides for agricultural use.

PubMed: 39621770
DOI: 10.1371/journal.ppat.1012745

実験手法

SOLUTION NMR