A Potent Anti-SpuE Antibody Allosterically Inhibits Type III Secretion System and Attenuates Virulence of Pseudomonas Aeruginosa

Zhang, Yang; Sun, Xiaomin; Qian, Yang‐Yang; Yi, Hongfei; Song, Ke; Zhu, Huanhu; Zonta, Francesco; Chen, Weizhong; Ji, Quanjiang; Miersch, Shane; Sidhu, Sachdev S.; Wu, Donghui

doi:10.1016/j.jmb.2019.10.026

Cited by 12 publications

(9 citation statements)

References 69 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The more common sequence variants in particular clustered in the domain that determines the binding specificity for biogenic amines. Considering the fact that the SpuE-mediated uptake of spermidine from the mammalian host leads to transcriptional upregulation of the type III secretion system [45, 46], the major virulence determinant of P. aeruginosa , it is reasonable to assume that within the scenario of host–pathogen interactions the sequence diversity of SpuE is translated into a batch of functional variants of differential pathogenicity.…”

Section: Resultsmentioning

confidence: 99%

Sequence diversity of the Pseudomonas aeruginosa population in loci that undergo microevolution in cystic fibrosis airways

Fischer

Klockgether

Sorribes

et al. 2021

Access Microbiology

View full text Add to dashboard Cite

Five hundred and thirty-four unrelated Pseudomonas aeruginosa isolates from inanimate habitats, patients with cystic fibrosis (CF) and other human infections were sequenced in 19 genes that had been identified previously as the hot spots of genomic within-host evolution in serial isolates from 12 CF lungs. Amplicon sequencing confirmed a significantly higher sequence diversity of the 19 loci in P. aeruginosa isolates from CF patients compared to those from other habitats, but this overrepresentation was mainly due to the larger share of synonymous substitutions. Correspondingly, non-synonymous substitutions were either rare (gltT, lepA, ptsP) or benign (nuoL, fleR, pelF) in some loci. Other loci, however, showed an accumulation of non-neutral coding variants. Strains from the CF habitat were often mutated at evolutionarily conserved positions in the elements of stringent response (RelA, SpoT), LPS (PagL), polyamine transport (SpuE, SpuF) and alginate biosynthesis (AlgG, AlgU). The strongest skew towards the CF lung habitat was seen for amino acid sequence variants in AlgG that clustered in the carbohydrate-binding/sugar hydrolysis domain. The master regulators of quorum sensing lasR and rhlR were frequent targets for coding variants in isolates from chronic and acute human infections. Unique variants in lasR showed strong evidence of positive selection indicated by d N/d S values of ~4. The pelA gene that encodes a multidomain enzyme involved in both the formation and dispersion of Pel biofilms carried the highest number of single-nucleotide variants among the 19 genes and was the only gene with a higher frequency of missense mutations in P. aeruginosa strains from non-CF habitats than in isolates from CF airways. PelA protein variants are widely distributed in the P. aeruginosa population. In conclusion, coding variants in a subset of the examined loci are indeed characteristic for the adaptation of P. aeruginosa to the CF airways, but for other loci the elevated mutation rate is more indicative of infections in human habitats (lasR, rhlR) or global diversifying selection (pelA).

show abstract

Section: Resultsmentioning

confidence: 99%

Sequence diversity of the Pseudomonas aeruginosa population in loci that undergo microevolution in cystic fibrosis airways

Fischer

Klockgether

Sorribes

et al. 2021

Access Microbiology

View full text Add to dashboard Cite

show abstract

“…Multidrug-resistant gram-negative bacteria have a significant impact on public health, and SPD uptake has been linked to the expression of type III secretion (T3SS) system genes ( 23 ), which are an essential part in their pathogenesis ( 24 , 25 ). Therefore, a SpuE antibody was designed to prevent SPD transport and ultimately weakening Pseudomonas aeruginosa infection ( 26 ). Bacterial pathogens have proven their adaptability numerous times, and by having promiscuous PBPs like PotF (or its homologs) at their disposal, they might just be able to hijack another system to facilitate SPD uptake and bypass treatment methods in the long run.…”

Section: Discussionmentioning

confidence: 99%

Fine-tuning spermidine binding modes in the putrescine binding protein PotF

Kröger

Shanmugaratnam

Scheib

et al. 2021

Journal of Biological Chemistry

View full text Add to dashboard Cite

A profound understanding of the molecular interactions between receptors and ligands is important throughout diverse research, such as protein design, drug discovery, or neuroscience. What determines specificity and how do proteins discriminate against similar ligands? In this study, we analyzed factors that determine binding in two homologs belonging to the well-known superfamily of periplasmic binding proteins, PotF and PotD. Building on a previously designed construct, modes of polyamine binding were swapped. This change of specificity was approached by analyzing local differences in the binding pocket as well as overall conformational changes in the protein. Throughout the study, protein variants were generated and characterized structurally and thermodynamically, leading to a specificity swap and improvement in affinity. This dataset not only enriches our knowledge applicable to rational protein design but also our results can further lay groundwork for engineering of specific biosensors as well as help to explain the adaptability of pathogenic bacteria.

show abstract

“…SpuE is a transcriptional regulator of the T3SS in P. aeruginosa and regulates the expression of ExsA, a master regulator of the T3SS [2], via inhibition of the exsCEBA promoter. Zhang et al derived an anti-SpuE nanobody fused with a membrane-penetrating peptide (scFv5-MPP) to assist in delivering the nanobody into the bacterial cell [145]. scFv5-MPP allosterically inhibits the expression of the T3SS and attenuates virulence of P. aeruginosa in a C. elegans animal gut infection model.…”

Section: Antibodies Targeting Intracellular Effectors and Transcriptimentioning

confidence: 99%

“…X-ray crystallography and molecular dynamics simulations were used to observe conformational changes upon antibody binding to SpuE. This conformational change may cause a reduction in spermidine uptake by P. aeruginosa leading to attenuation of virulence similar to physical neutralization [145]. A similar antibody, Mab 4E4, protects A549 cells against P. aeruginosa infection by reducing T3SS expression and polyamine uptake.…”

Section: Antibodies Targeting Intracellular Effectors and Transcriptimentioning

confidence: 99%

Antibodies Inhibiting the Type III Secretion System of Gram-Negative Pathogenic Bacteria

Hotinger

May

2020

Antibodies

View full text Add to dashboard Cite

Pathogenic bacteria are a global health threat, with over 2 million infections caused by Gram-negative bacteria every year in the United States. This problem is exacerbated by the increase in resistance to common antibiotics that are routinely used to treat these infections, creating an urgent need for innovative ways to treat and prevent virulence caused by these pathogens. Many Gram-negative pathogenic bacteria use a type III secretion system (T3SS) to inject toxins and other effector proteins directly into host cells. The T3SS has become a popular anti-virulence target because it is required for pathogenesis and knockouts have attenuated virulence. It is also not required for survival, which should result in less selective pressure for resistance formation against T3SS inhibitors. In this review, we will highlight selected examples of direct antibody immunizations and the use of antibodies in immunotherapy treatments that target the bacterial T3SS. These examples include antibodies targeting the T3SS of Pseudomonas aeruginosa, Yersinia pestis, Escherichia coli, Salmonella enterica, Shigella spp., and Chlamydia trachomatis.

show abstract

A Potent Anti-SpuE Antibody Allosterically Inhibits Type III Secretion System and Attenuates Virulence of Pseudomonas Aeruginosa

Cited by 12 publications

References 69 publications

Sequence diversity of the Pseudomonas aeruginosa population in loci that undergo microevolution in cystic fibrosis airways

Sequence diversity of the Pseudomonas aeruginosa population in loci that undergo microevolution in cystic fibrosis airways

Fine-tuning spermidine binding modes in the putrescine binding protein PotF

Antibodies Inhibiting the Type III Secretion System of Gram-Negative Pathogenic Bacteria

Contact Info

Product

Resources

About