Substrate access mechanism in a novel membrane-bound phospholipase A of <i>Pseudomonas aeruginosa</i> concordant with specificity and regioselectivity

Ahmad, Sameena; Strunk, Christoph Heinrich; Schott‐Verdugo, Stephan; Jaeger, Karl‐Erich; Kovačić, Filip; Gohlke, Holger

doi:10.1101/2021.06.29.450291

Cited by 3 publications

(3 citation statements)

References 117 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…49 Within these stages, the trajectory closest to the Jarzynski average is determined, and the final state of this trajectory is used as starting point for the consecutive stage. 50 Thus, trajectories contributing little to the overall PMF are disregarded in the next stage, that way reducing the total number of simulations to be performed.…”

Section: Methodsmentioning

confidence: 99%

Rational Design Yields Molecular Insights on Leaf-Binding of Anchor Peptides

Dittrich

Brethauer

Goncharenko

et al. 2022

ACS Appl. Mater. Interfaces

Self Cite

View full text Add to dashboard Cite

In times of a constantly growing world population and increasing demand for food, sustainable agriculture is crucial. The rainfastness of plant protection agents is of pivotal importance to reduce the amount of applied nutrients, herbicides, and fungicides. As a result of protective agent wash-off, plant protection is lost, and soils and groundwater are severely polluted. To date, rainfastness of plant protection products has been achieved by adding polymeric adjuvants to the agrochemicals. However, polymeric adjuvants will be regarded as microplastics in the future, and environmentally friendly alternatives are needed. Anchor peptides (APs) are promising biobased and biodegradable adhesion promoters. Although the adhesion of anchor peptides to artificial surfaces, such as polymers, has already been investigated in theory and experimentally, exploiting the adhesion to biological surfaces remains challenging. The complex nature and composition of biological surfaces such as plant leaves and fruit surfaces complicate the generation of accurate models. Here, we present the first detailed three-layered atomistic model of the surface of apple leaves and use it to compute free energy profiles of the adhesion and desorption of APs to and from that surface. Our model is validated by a novel fluorescence-based microtiter plate (MTP) assay that mimics these complex processes and allows for quantifying them. For the AP Macaque Histatin, we demonstrate that aromatic and positively charged amino acids are essential for binding to the waxy apple leaf surface. The established protocols should generally be applicable for tailoring the binding properties of APs to biological interfaces.

show abstract

Section: Methodsmentioning

confidence: 99%

Rational Design Yields Molecular Insights on Leaf-Binding of Anchor Peptides

Dittrich

Brethauer

Goncharenko

et al. 2022

ACS Appl. Mater. Interfaces

Self Cite

View full text Add to dashboard Cite

show abstract

“…Molecular dynamics (MD simulations) can fill this gap, by providing realistic insights into the interactions of proteins with model membranes (28)(29)(30). In particular, MD simulations have been used to predict membrane-binding interfaces, identify lipid binding sites and membrane-inserting regions for key signaling proteins including PTEN, KRAS, bacterial phospholipases, and pleckstrin homology domains (31)(32)(33)(34)(35).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the membrane interactions of phospholipase Cγ reveals key features of the active enzyme

et al. 2022

View full text Add to dashboard Cite

PLCγ enzymes are autoinhibited in resting cells and form key components of intracellular signaling that are also linked to disease development. Insights into physiological and aberrant activation of PLCγ require understanding of an active, membrane-bound form, which can hydrolyze inositol-lipid substrates. Here, we demonstrate that PLCγ1 cannot bind membranes unless the autoinhibition is disrupted. Through extensive molecular dynamics simulations and experimental evidence, we characterize membrane binding by the catalytic core domains and reveal previously unknown sites of lipid interaction. The identified sites act in synergy, overlap with autoinhibitory interfaces, and are shown to be critical for the phospholipase activity in cells. This work provides direct evidence that PLCγ1 is inhibited through obstruction of its membrane-binding surfaces by the regulatory region and that activation must shift PLCγ1 to a conformation competent for membrane binding. Knowledge of the critical sites of membrane interaction extends the mechanistic framework for activation, dysregulation, and therapeutic intervention.

show abstract

“…A crystal structure and biochemical and computational results provided a mechanism of how dimer to monomer transition of PlaF and subsequent rotation of PlaF at the membrane surface leads to recognition of PL substrate, its extraction from the membrane through the tunnel to the active site where it is hydrolysed and the products are likely released to the membrane [38, 41, 42]. We suggest that PlaF-dependent modulation of the membrane PL composition is responsible for the strongly attenuated virulence of P. aeruginosa ΔplaF in a Galleria mellonella virulence assay and macrophage cytotoxicity assay.…”

Section: Introductionmentioning

confidence: 99%

Virulence adaptation ofPseudomonas aeruginosaphospholipase mutant with altered membrane phospholipid composition

Caliskan

Poschmann

Gudzuhn

et al. 2022

Preprint

View full text Add to dashboard Cite

Membrane protein and phospholipid (PL) composition changes in response to environmental cues and during infections. Covalent modification and remodelling of the acyl chain length of PLs is an important bacterial adaptation mechanism. However, little is known about which bacterial pathways are regulated in response to altered PL composition. Here, we showed that P. aeruginosa phospholipase A, PlaF, which modulates membrane PL composition, is important for biofilm biogenesis, and we performed whole-cell quantitative proteomics of P. aeruginosa wild-type and delta plaF biofilms to identify pathways regulated by PlaF. The results revealed profound alterations in the abundance of several two-component systems (TCSs), including accumulation of PprAB, which controls the transition to biofilm. Furthermore, a unique phosphorylation pattern of transcriptional regulators, transporters and metabolic enzymes, as well as differential production of seven proteases, in delta plaF, indicate that PlaF-mediated virulence adaptation involves complex transcriptional and posttranscriptional regulation. Moreover, proteomics revealed the depletion of pyoverdine-mediated iron uptake pathway proteins in delta plaF, which agrees with the decreased concentrations of extracellular pyoverdine and intracellular iron and is likely responsible for its prolonged lag growth phase, presumably due to reduced iron uptake. Conversely, the accumulation of proteins from alternative iron-uptake systems in delta plaF suggests that PlaF may function as a switch between different iron-acquisition pathways. The observation that ∆plaF accumulates PL-acyl chain modifying and PL synthesis enzymes reveals novel insights into the role of PlaF for membrane PL homeostasis. Although the precise mechanism by which PlaF simultaneously affects multiple pathways remains to be elucidated, we suggest that PlaF-catalyses the degradation of PLs which then serve as a signal that is amplified by proteins of two-component, phosphorylation and proteolytic degradation systems to elicit the global adaptive response in P. aeruginosa.

show abstract

Substrate access mechanism in a novel membrane-bound phospholipase A of Pseudomonas aeruginosa concordant with specificity and regioselectivity

Cited by 3 publications

References 117 publications

Rational Design Yields Molecular Insights on Leaf-Binding of Anchor Peptides

Rational Design Yields Molecular Insights on Leaf-Binding of Anchor Peptides

Characterization of the membrane interactions of phospholipase Cγ reveals key features of the active enzyme

Virulence adaptation ofPseudomonas aeruginosaphospholipase mutant with altered membrane phospholipid composition

Contact Info

Product

Resources

About