Effect of DMSO on the Mechanical and Structural Properties of Model and Biological Membranes

Gironi, Beatrice; Kahveci, Zehra; McGill, Beth; Lechner, Bob‐Dan; Pagliara, Stefano; Metz, Jeremy; Morresi, Assunta; Palombo, Francesca; Sassi, Paola; Petrov, Peter G.

doi:10.1016/j.bpj.2020.05.037

Cited by 52 publications

(49 citation statements)

References 68 publications

(109 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Due to the weak solubility of LF11-324 in buffer, AMP stock solutions were prepared by adding acetic acid and DMSO, up to 0.3% and 3% vol/vol, respectively. Prior to each measurement, the concentrations of such compounds were systematically lowered down to 0.01% acetic acid and 0.1% vol/vol DMSO (final pH 7.2), in order to minimize, or even remove, any effect on the membrane structure ( 29 ). AMP concentrations of the stock solutions were determined by comparing against the absorption band of the aromatic amino acids with the spectrophotometer NanoDrop ND-1000 (Thermo Fisher Scientific, Waltham, MA).…”

Section: Methodsmentioning

confidence: 99%

Bridging the Antimicrobial Activity of Two Lactoferricin Derivatives in E. coli and Lipid-Only Membranes

Marx

Semeraro

Mandl

et al. 2021

Front. Med. Technol.

View full text Add to dashboard Cite

We coupled the antimicrobial activity of two well-studied lactoferricin derivatives, LF11-215 and LF11-324, in Escherichia coli and different lipid-only mimics of its cytoplasmic membrane using a common thermodynamic framework for peptide partitioning. In particular, we combined an improved analysis of microdilution assays with ζ-potential measurements, which allowed us to discriminate between the maximum number of surface-adsorbed peptides and peptides fully partitioned into the bacteria. At the same time, we measured the partitioning of the peptides into vesicles composed of phosphatidylethanolamine (PE), phosphatidylgylcerol (PG), and cardiolipin (CL) mixtures using tryptophan fluorescence and determined their membrane activity using a dye leakage assay and small-angle X-ray scattering. We found that the vast majority of LF11-215 and LF11-324 readily enter inner bacterial compartments, whereas only 1−5% remain surface bound. We observed comparable membrane binding of both peptides in membrane mimics containing PE and different molar ratios of PG and CL. The peptides' activity caused a concentration-dependent dye leakage in all studied membrane mimics; however, it also led to the formation of large aggregates, part of which contained collapsed multibilayers with sandwiched peptides in the interstitial space between membranes. This effect was least pronounced in pure PG vesicles, requiring also the highest peptide concentration to induce membrane permeabilization. In PE-containing systems, we additionally observed an effective shielding of the fluorescent dyes from leakage even at highest peptide concentrations, suggesting a coupling of the peptide activity to vesicle fusion, being mediated by the intrinsic lipid curvatures of PE and CL. Our results thus show that LF11-215 and LF11-324 effectively target inner bacterial components, while the stored elastic stress makes membranes more vulnerable to peptide translocation.

show abstract

Section: Methodsmentioning

confidence: 99%

Bridging the Antimicrobial Activity of Two Lactoferricin Derivatives in E. coli and Lipid-Only Membranes

Marx

Semeraro

Mandl

et al. 2021

Front. Med. Technol.

View full text Add to dashboard Cite

show abstract

“…DMSO is found in many natural sources including fruit ( Pearson et al 1981 ) and marine algae ( Lee and Mora 1999 ). In addition to its laboratory use as a solvent and cryoprotectant due to effects on cellular membranes ( Notman et al 2006 ; Gurtovenko and Anwar 2007 ; Gironi et al 2020 ), DMSO also affects protein folding and stability in vitro ( Rammler and Zaffaroni 1967 ). Like GdnHCl, DMSO (at 2.5% v/v) cures yeast cells of at least one prion ( Tuite et al 1981 ).…”

Section: Resultsmentioning

confidence: 99%

Chemical rescue of mutant proteins in living Saccharomyces cerevisiae cells by naturally occurring small molecules

Hassell

Steingesser

Denney

et al. 2021

G3 Genes|Genomes|Genetics

View full text Add to dashboard Cite

Intracellular proteins function in a complex milieu wherein small molecules influence protein folding and act as essential cofactors for enzymatic reactions. Thus protein function depends not only on amino acid sequence but also on the concentrations of such molecules, which are subject to wide variation between organisms, metabolic states, and environmental conditions. We previously found evidence that exogenous guanidine reverses the phenotypes of specific budding yeast septin mutants by binding to a WT septin at the former site of an Arg side chain that was lost during fungal evolution. Here we used a combination of targeted and unbiased approaches to look for other cases of “chemical rescue” by naturally occurring small molecules. We report in vivo rescue of hundreds of Saccharomyces cerevisiae mutants representing a variety of genes, including likely examples of Arg or Lys side chain replacement by the guanidinium ion. Failed rescue of targeted mutants highlight features required for rescue, as well as key differences between the in vitro and in vivo environments. Some non-Arg mutants rescued by guanidine likely result from “off-target” effects on specific cellular processes in WT cells. Molecules isosteric to guanidine and known to influence protein folding had a range of effects, from essentially none for urea, to rescue of a few mutants by DMSO. Strikingly, the osmolyte trimethylamine-N-oxide rescued ∼20% of the mutants we tested, likely reflecting combinations of direct and indirect effects on mutant protein function. Our findings illustrate the potential of natural small molecules as therapeutic interventions and drivers of evolution.

show abstract

“…In the laboratory setting DMSO is widely used as a solvent and cryoprotectant due to effects on cellular membranes, which vary with membrane lipid composition and DMSO concentration but include thinning, stiffening, softening, and probably formation of pores (101–103). DMSO also affects protein folding and stability in vitro (104).…”

Section: Resultsmentioning

confidence: 99%

Chemical rescue of mutant proteins in living cells by naturally occurring small molecules

Hassell

Steingesser

Denney

et al. 2021

Preprint

View full text Add to dashboard Cite

Intracellular proteins function in a complex milieu wherein small molecules influence protein folding and act as essential cofactors for enzymatic reactions. Thus protein function depends not only on amino acid sequence but also on the concentrations of such molecules, which are subject to wide variation between organisms, metabolic states, and environmental conditions. We previously found evidence that exogenous guanidine reverses the phenotypes of specific budding yeast septin mutants by binding to a WT septin at the former site of an Arg side chain that was lost during fungal evolution. Here we used a combination of targeted and unbiased approaches to look for other cases of chemical rescue by naturally occurring small molecules. We report in vivo rescue of hundreds of yeast mutants representing a variety of genes, including likely examples of Arg or Lys side chain replacement by the guanidinium ion. Failed rescue of targeted mutants highlight features required for rescue, as well as key differences between the in vitro and in vivo environments. Some non-Arg mutants rescued by guanidine likely result from off-target effects on specific cellular processes in WT cells. Molecules isosteric to guanidine and known to influence protein folding had a range of effects, from essentially none for urea, to rescue of a few mutants by DMSO. Strikingly, the osmolyte trimethylamine-N-oxide rescued ~20% of the mutants we tested, likely reflecting combinations of direct and indirect effects on mutant protein function. Our findings illustrate the potential of natural small molecules as therapeutic interventions and drivers of evolution.

show abstract

Effect of DMSO on the Mechanical and Structural Properties of Model and Biological Membranes

Cited by 52 publications

References 68 publications

Bridging the Antimicrobial Activity of Two Lactoferricin Derivatives in E. coli and Lipid-Only Membranes

Bridging the Antimicrobial Activity of Two Lactoferricin Derivatives in E. coli and Lipid-Only Membranes

Chemical rescue of mutant proteins in living Saccharomyces cerevisiae cells by naturally occurring small molecules

Chemical rescue of mutant proteins in living cells by naturally occurring small molecules

Contact Info

Product

Resources

About