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.

doi:10.1074/jbc.m111.311316

Cited by 80 publications

(70 citation statements)

References 28 publications

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“…It has been suggested that this chemical and structural diversity impacts peptide activity, as found in other families of small disulfide-linked peptides that exhibit rich chemical and structural diversity (4,49). Indeed, experiments on modified versions of the NCR247 peptide indicated that the presence and order of disulfide bonds between cysteine residues significantly influences peptide antimicrobial activity (50). It has been proposed that NCR peptides (i) alter the S. meliloti cell cycle and metabolism to achieve a more efficient symbiosis (4), (ii) sanction non-nitrogen-fixing rhizobia (4), (iii) kill senescent bacteroids so that their nutrients can be absorbed by the plant (4,48), and (iv) serve as signals to the plant (48).…”

Section: Discussionmentioning

confidence: 99%

Host plant peptides elicit a transcriptional response to control theSinorhizobium meliloticell cycle during symbiosis

Penterman

Abo

Nisco

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The α-proteobacterium Sinorhizobium meliloti establishes a chronic intracellular infection during the symbiosis with its legume hosts. Within specialized host cells, S. meliloti differentiates into highly polyploid, enlarged nitrogen-fixing bacteroids. This differentiation is driven by host cells through the production of defensin-like peptides called "nodule-specific cysteine-rich" (NCR) peptides. Recent research has shown that synthesized NCR peptides exhibit antimicrobial activity at high concentrations but cause bacterial endoreduplication at sublethal concentrations. We leveraged synchronized S. meliloti populations to determine how treatment with a sublethal NCR peptide affects the cell cycle and physiology of bacteria at the molecular level. We found that at sublethal levels a representative NCR peptide specifically blocks cell division and antagonizes Z-ring function. Gene-expression profiling revealed that the cell division block was produced, in part, through the substantial transcriptional response elicited by sublethal NCR treatment that affected ∼15% of the genome. Expression of critical cell-cycle regulators, including ctrA, and cell division genes, including genes required for Z-ring function, were greatly attenuated in NCR-treated cells. In addition, our experiments identified important symbiosis functions and stress responses that are induced by sublethal levels of NCR peptides and other antimicrobial peptides. Several of these stress-response pathways also are found in related α-proteobacterial pathogens and might be used by S. meliloti to sense host cues during infection. Our data suggest a model in which, in addition to provoking stress responses, NCR peptides target intracellular regulatory pathways to drive S. meliloti endoreduplication and differentiation during symbiosis.rhizobia-legume | host-microbe interactions

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Section: Discussionmentioning

confidence: 99%

Host plant peptides elicit a transcriptional response to control theSinorhizobium meliloticell cycle during symbiosis

Penterman

Abo

Nisco

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…Cys is a very important metabolite not only as a proteinogenic amino acid, but also by its function as a precursor of essential biomolecules. In proteins, the thiol group of Cys, susceptible of oxidation to render disulfide bridges, stabilizes the tertiary and quaternary protein structures and, consequently, determines protein function (Haag et al 2012 ), and often located in enzyme active sites are essential for catalysis (Richau et al 2012 ). Moreover, the reversible conversion between free thiols groups and disulfide bridges is the basis of protein redox regulation (Buchanan and Balmer 2005 ).…”

Section: Introductionmentioning

confidence: 99%

Signaling in the plant cytosol: cysteine or sulfide?

et al. 2014

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Cysteine (Cys) is the first organic compound containing reduced sulfur that is synthesized in the last stage of plant photosynthetic assimilation of sulfate. It is a very important metabolite not only because it is crucial for the structure, function and regulation of proteins but also because it is the precursor molecule of an enormous number of sulfur-containing metabolites essential for plant health and development. The biosynthesis of Cys is accomplished by the sequential reaction of serine acetyltransferase (SAT) and O-acetylserine(thiol)synthase (OASTL). In Arabidopsis thaliana, the analysis of specific mutants of members of the SAT and OASTL families has demonstrated that the cytosol is the compartment where the bulk of Cys synthesis takes place and that the cytosolic OASTL enzyme OAS-A1 is the responsible enzyme. Another member of the OASTL family is DES1, a novel L-cysteine desulfhydrase that catalyzes the desulfuration of Cys to produce sulfide, thus acting in a manner opposite to that of OAS-A1. Detailed studies of the oas-a1 and

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“…Its disulfide bridge plays important roles in its biological activities. Similarly, Haag et al (19) found that the nodule-specific cysteinerich peptide (NCR247) had decreased activity against Sinorhizobium meliloti when cysteines were substituted for serines or the S-S bridges were changed from cysteines 1-2 and 3-4 to cysteines 1-3 and 2-4.…”

Section: Discussionmentioning

confidence: 99%

The Disulfide Bond of the Peptide Thanatin Is Dispensible for Its Antimicrobial Activity In Vivo and In Vitro

Niu

Zhou

et al. 2016

Antimicrob Agents Chemother

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b Thanatin (THA) displays potent antibiotic activity, especially against extended-spectrum-␤-lactamase (ESBL)-producing Escherichia coli both in vitro and in vivo, with minimal hemolytic toxicity and satisfactory stability in plasma. However, the high cost of thanatin significantly limits its development and clinical application. To reduce the cost of peptide synthesis, a formulation of cyclic thanatin (C-thanatin) called linear thanatin (L-thanatin) was synthesized and its activity was evaluated in vivo and in vitro. Results showed that C-thanatin and L-thanatin MICs did not differ against eight Gram-negative and two Gram-positive bacterial strains. Furthermore, the survival rates of ESBL-producing-E. coli-infected mice were consistent after C-thanatin or L-thanatin treatment at 5 or 10 mg/kg of body weight. Neither C-thanatin nor L-thanatin showed toxicity for human red blood cells (hRBCs) and human umbilical vein endothelial cells (HUVECs) at a concentration as high as 256 g/ml. Results of circular dichroism spectroscopy indicated that the secondary structure of L-thanatin is extremely similar to that of C-thanatin. Membrane permeabilization and depolarization assays showed that C-thanatin and L-thanatin have similar abilities to permeabilize the outer and inner membranes and to induce membrane depolarization in ESBL-producing E. coli. However, neither of them caused significant HUVEC membrane permeability. These findings indicate that the two peptides have similar effects on bacterial cell membranes and that the disulfide bond in thanatin is not essential for its antimicrobial activities in vivo and in vitro. L-thanatin is thus a promising low-cost peptide candidate for treating ESBL-producing E. coli infections. T hanatin (THA) is a cationic antimicrobial peptide that was first discovered in the hemipteran insect Podisus maculiventris.This peptide is composed of 21 amino acids (GSKKPVPIIYCNR RTGKCQRM), with an intramolecular disulfide bridge between the two cysteines at positions 11 and 18. Our previous studies revealed that thanatin exhibits potent antibiotic activity, especially against extended-spectrum-␤-lactamase (ESBL)-producing Esc herichia coli, both in vitro and in vivo, with a low inherent ability to induce microbial resistance, minimal hemolytic toxicity, and high stability in plasma (1, 2).Despite their promise for clinical benefit, the high cost of producing peptide drugs such as thanatin significantly limits their development and clinical application. Compared with conventional antibiotics, the costs of cationic antimicrobial peptides are always prohibitively high for large-scale production (3). Intrachain disulfide bonds exist in many naturally occurring peptides and play important roles in biological activities. However, synthesis of peptide drugs with one or more disulfide bridges is complicated and expensive (4).Previous studies found that the disulfide bonds of protegrin-1 are not essential for its antimicrobial and hemolytic activities in vitro (5). To reduce the cost of thanatin synthes...

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Role of Cysteine Residues and Disulfide Bonds in the Activity of a Legume Root Nodule-specific, Cysteine-rich Peptide

Cited by 80 publications

References 28 publications

Host plant peptides elicit a transcriptional response to control theSinorhizobium meliloticell cycle during symbiosis

Host plant peptides elicit a transcriptional response to control theSinorhizobium meliloticell cycle during symbiosis

Signaling in the plant cytosol: cysteine or sulfide?

The Disulfide Bond of the Peptide Thanatin Is Dispensible for Its Antimicrobial Activity In Vivo and In Vitro

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