Role of cysteine residues and disulfide bonds in the activity of a legume root nodule-specific, cysteine-rich peptide

Andreas F Haag, Bernhard Kerscher, Sergio Dall'Angelo, Monica Sani, Renato Longhi, Mikhail Baloban, Heather M Wilson, Peter Mergaert, Matteo Zanda, Gail P Ferguson

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

The root nodules of certain legumes including Medicago truncatula produce >300 different nodule-specific cysteine-rich (NCR) peptides. Medicago NCR antimicrobial peptides (AMPs) mediate the differentiation of the bacterium, Sinorhizobium meliloti into a nitrogen-fixing bacteroid within the legume root nodules. In vitro, NCR AMPs such as NCR247 induced bacteroid features and exhibited antimicrobial activity against S. meliloti. The bacterial BacA protein is critical to prevent S. meliloti from being hypersensitive toward NCR AMPs. NCR AMPs are cationic and have conserved cysteine residues, which form disulfide (S–S) bridges. However, the natural configuration of NCR AMP S–S bridges and the role of these in the activity of the peptide are unknown. In this study, we found that either cysteine replacements or S–S bond modifications influenced the activity of NCR247 against S. meliloti. Specifically, either substitution of cysteines for serines, changing the S–S bridges from cysteines 1–2, 3–4 to 1–3, 2–4 or oxidation of NCR247 lowered its activity against S. meliloti. We also determined that BacA specifically protected S. meliloti against oxidized NCR247. Due to the large number of different NCRs synthesized by legume root nodules and the importance of bacterial BacA proteins for prolonged host infections, these findings have important implications for analyzing the function of these novel peptides and the protective role of BacA in the bacterial response toward these peptides.
Original languageEnglish
Pages (from-to)10791-10798
Number of pages8
JournalThe Journal of Biological Chemistry
Volume287
Issue number14
Early online date17 Feb 2012
DOIs
Publication statusPublished - 30 Mar 2012

Fingerprint

Fabaceae
Disulfides
Cysteine
Sinorhizobium meliloti
Peptides
Bacterial Proteins
Antimicrobial Cationic Peptides
Medicago truncatula
Medicago
Serine
Bacteria
Substitution reactions
Nitrogen
Oxidation
Infection

Keywords

  • antimicrobial peptides
  • bacteria
  • microbiology
  • peptide conformation
  • symbiosis
  • BacA
  • defensins
  • nodule-specific, cysteine-rich peptides
  • rhizobia
  • Sinorhizobium meliloti

Cite this

Role of cysteine residues and disulfide bonds in the activity of a legume root nodule-specific, cysteine-rich peptide. / Haag, Andreas F; Kerscher, Bernhard; Dall'Angelo, Sergio; Sani, Monica; Longhi, Renato; Baloban, Mikhail; Wilson, Heather M; Mergaert, Peter; Zanda, Matteo; Ferguson, Gail P.

In: The Journal of Biological Chemistry, Vol. 287, No. 14, 30.03.2012, p. 10791-10798.

Research output: Contribution to journalArticle

Haag, Andreas F ; Kerscher, Bernhard ; Dall'Angelo, Sergio ; Sani, Monica ; Longhi, Renato ; Baloban, Mikhail ; Wilson, Heather M ; Mergaert, Peter ; Zanda, Matteo ; Ferguson, Gail P. / Role of cysteine residues and disulfide bonds in the activity of a legume root nodule-specific, cysteine-rich peptide. In: The Journal of Biological Chemistry. 2012 ; Vol. 287, No. 14. pp. 10791-10798.
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T1 - Role of cysteine residues and disulfide bonds in the activity of a legume root nodule-specific, cysteine-rich peptide

AU - Haag, Andreas F

AU - Kerscher, Bernhard

AU - Dall'Angelo, Sergio

AU - Sani, Monica

AU - Longhi, Renato

AU - Baloban, Mikhail

AU - Wilson, Heather M

AU - Mergaert, Peter

AU - Zanda, Matteo

AU - Ferguson, Gail P

PY - 2012/3/30

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N2 - The root nodules of certain legumes including Medicago truncatula produce >300 different nodule-specific cysteine-rich (NCR) peptides. Medicago NCR antimicrobial peptides (AMPs) mediate the differentiation of the bacterium, Sinorhizobium meliloti into a nitrogen-fixing bacteroid within the legume root nodules. In vitro, NCR AMPs such as NCR247 induced bacteroid features and exhibited antimicrobial activity against S. meliloti. The bacterial BacA protein is critical to prevent S. meliloti from being hypersensitive toward NCR AMPs. NCR AMPs are cationic and have conserved cysteine residues, which form disulfide (S–S) bridges. However, the natural configuration of NCR AMP S–S bridges and the role of these in the activity of the peptide are unknown. In this study, we found that either cysteine replacements or S–S bond modifications influenced the activity of NCR247 against S. meliloti. Specifically, either substitution of cysteines for serines, changing the S–S bridges from cysteines 1–2, 3–4 to 1–3, 2–4 or oxidation of NCR247 lowered its activity against S. meliloti. We also determined that BacA specifically protected S. meliloti against oxidized NCR247. Due to the large number of different NCRs synthesized by legume root nodules and the importance of bacterial BacA proteins for prolonged host infections, these findings have important implications for analyzing the function of these novel peptides and the protective role of BacA in the bacterial response toward these peptides.

AB - The root nodules of certain legumes including Medicago truncatula produce >300 different nodule-specific cysteine-rich (NCR) peptides. Medicago NCR antimicrobial peptides (AMPs) mediate the differentiation of the bacterium, Sinorhizobium meliloti into a nitrogen-fixing bacteroid within the legume root nodules. In vitro, NCR AMPs such as NCR247 induced bacteroid features and exhibited antimicrobial activity against S. meliloti. The bacterial BacA protein is critical to prevent S. meliloti from being hypersensitive toward NCR AMPs. NCR AMPs are cationic and have conserved cysteine residues, which form disulfide (S–S) bridges. However, the natural configuration of NCR AMP S–S bridges and the role of these in the activity of the peptide are unknown. In this study, we found that either cysteine replacements or S–S bond modifications influenced the activity of NCR247 against S. meliloti. Specifically, either substitution of cysteines for serines, changing the S–S bridges from cysteines 1–2, 3–4 to 1–3, 2–4 or oxidation of NCR247 lowered its activity against S. meliloti. We also determined that BacA specifically protected S. meliloti against oxidized NCR247. Due to the large number of different NCRs synthesized by legume root nodules and the importance of bacterial BacA proteins for prolonged host infections, these findings have important implications for analyzing the function of these novel peptides and the protective role of BacA in the bacterial response toward these peptides.

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KW - bacteria

KW - microbiology

KW - peptide conformation

KW - symbiosis

KW - BacA

KW - defensins

KW - nodule-specific, cysteine-rich peptides

KW - rhizobia

KW - Sinorhizobium meliloti

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JF - The Journal of Biological Chemistry

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