Binding of doxorubicin to cardiolipin as compared to other anionic phospholipids—An evaluation of electrostatic effects

1991 ◽  
Vol 11 (5) ◽  
pp. 275-284 ◽  
Author(s):  
Frits A. de Wolf
Keyword(s):  
Drug Concentration ◽  
Membrane Surface ◽  
Local Concentration ◽  
Lipid Phosphorus ◽  
Apparent Discrepancy ◽  
Electrostatic Effects ◽  
Anionic Phospholipids ◽  
Large Unilamellar Vesicles ◽  
Free Doxorubicin ◽  
Binding Level

The binding of doxorubicin to large unilamellar vesicles consisting of cardiolipin or other anionic phospholipids was analyzed in terms of the local drug concentration at the membrane surface, according to the Gouy-Chapman theory. The analysis suggests strong positive binding cooperativity. Part of the drug binds in the uncharged form. The affinity for cardiolipin and other anionic phospholipids is comparable. A binding level of 0.5 doxorubicin per lipid-phosphorus is reached when the local concentration of free doxorubicin monomer-equivalents at the membrane surface is about 0.2–0.7 mM. This contrasts with earlier findings indicating a 300–1000 fold higher affinity for cardiolipin. The present analysis provides an explanation for this apparent discrepancy.

scholarly journals Molecular determinants of KA1 domain-mediated autoinhibition and phospholipid activation of MARK1 kinase

2017 ◽  
Vol 474 (3) ◽  
pp. 385-398 ◽  
Author(s):  
Ryan P. Emptage ◽  
Mark A. Lemmon ◽  
Kathryn M. Ferguson
Keyword(s):  
Membrane Surface ◽  
Kinase Domain ◽  
Site Directed Mutagenesis ◽  
Anionic Phospholipid ◽  
Anionic Phospholipids ◽  
Disease States ◽  
C Terminus ◽  
Regulatory Module ◽  
Correct Location ◽  
Mechanistic Basis

Protein kinases are frequently regulated by intramolecular autoinhibitory interactions between protein modules that are reversed when these modules bind other ‘activating’ protein or membrane-bound targets. One group of kinases, the MAP/microtubule affinity-regulating kinases (MARKs) contain a poorly understood regulatory module, the KA1 (kinase associated-1) domain, at their C-terminus. KA1 domains from MARK1 and several related kinases from yeast to humans have been shown to bind membranes containing anionic phospholipids, and peptide ligands have also been reported. Deleting or mutating the C-terminal KA1 domain has been reported to activate the kinase in which it is found — also suggesting an intramolecular autoinhibitory role. Here, we show that the KA1 domain of human MARK1 interacts with, and inhibits, the MARK1 kinase domain. Using site-directed mutagenesis, we identify residues in the KA1 domain required for this autoinhibitory activity, and find that residues involved in autoinhibition and in anionic phospholipid binding are the same. We also demonstrate that a ‘mini’ MARK1 becomes activated upon association with vesicles containing anionic phospholipids, but only if the protein is targeted to these vesicles by a second signal. These studies provide a mechanistic basis for understanding how MARK1 and its relatives may require more than one signal at the membrane surface to control their activation at the correct location and time. MARK family kinases have been implicated in a plethora of disease states including Alzheimer's, cancer, and autism, so advancing our understanding of their regulatory mechanisms may ultimately have therapeutic value.

Channel Length Influence on the Performance of the Vacuum Membrane Distillation

2020 ◽  
Author(s):  
Abdulaziz M. Alasiri ◽  
Umar Alqsair ◽  
Sertac Cosman ◽  
Robert Krysko ◽  
Alparslan Oztekin
Keyword(s):  
Membrane Surface ◽  
Membrane Distillation ◽  
Channel Length ◽  
Water Vapor Transport ◽  
Membrane Thickness ◽  
Local Concentration ◽  
Permeate Flux ◽  
Hydrophobic Membrane ◽  
Vapor Flux ◽  
Temperature Polarization

Abstract The demand for freshwater has been increased globally. Membrane distillation (MD) technique can be an attractive option for desalination applications. MD is defined as a thermal-driven separation process that implements a hydrophobic membrane for allowing only water vapor transport through the membrane. VMD system is investigated in this study to examine its sensitivity toward the channel design. PTFE membrane is employed and treated as a functional surface where its main properties, such as porosity, tortuosity, pore diameter, and membrane thickness are defined. Different flow rates and inlet temperatures of the feed solution are involved to intensely study the effect of the channel length on VMD performance. The local concentration and temperature polarization coefficient and mass flux along the membrane surface are presented and discussed. With the increasing length of the module, concentration and temperature polarization levels are increased, and the vapor flux is decreased. It is shown that the permeate flux decreases linearly with the channel length. The slope of the permeate flux with length can be used to estimate the flux performance of modules with varying length.

scholarly journals Structure and Formation Mechanism of Antimicrobial Peptides Temporin B- and L-Induced Tubular Membrane Protrusion

2021 ◽  
Vol 22 (20) ◽  
pp. 11015
Author(s):  
Shan Zhang ◽  
Ming Ma ◽  
Zhuang Shao ◽  
Jincheng Zhang ◽  
Lei Fu ◽  
...  
Keyword(s):  
Antimicrobial Peptides ◽  
Morphological Changes ◽  
Membrane Surface ◽  
Coarse Grained ◽  
Tubular Structures ◽  
Membrane Pore ◽  
Anionic Phospholipids ◽  
Bactericidal Activities ◽  
Highly Hydrophobic ◽  
Dynamics Simulations

Temporins are a family of antimicrobial peptides (AMPs) isolated from frog skin, which are very short, weakly charged, and highly hydrophobic. They execute bactericidal activities in different ways from many other AMPs. This work investigated morphological changes of planar bilayer membranes composed of mixed zwitterionic and anionic phospholipids induced by temporin B and L (TB and TL) using all-atom and coarse-grained molecular dynamics simulations. We found that TB and TL fold to α-helices at the membrane surface and penetrate shallowly into the bilayer. These short AMPs have low propensity to induce membrane pore formation but possess high ability to extract lipids out. At relatively high peptide concentrations, the strong hydrophobicity of TB and TL promotes them to aggregate into clusters on the membrane surface. These aggregates attract a large amount of lipids out of the membrane to release compression induced by other dispersed peptides binding to the membrane. The extruded lipids mix evenly with the peptides in the cluster and form tubule-like protrusions. Certain water molecules follow the movement of lipids, which not only fill the cavities of the protrusion but also assist in maintaining the tubular structures. In contrast, the peptide-free leaflet remains intact. The present results unravel distinctive antimicrobial mechanisms of temporins disturbing membranes.

scholarly journals Role of anionic phospholipids in the adaptation of Bacillus subtilis to high salinity

Microbiology ◽  
2006 ◽  
Vol 152 (3) ◽  
pp. 605-616 ◽  
Author(s):  
Claudia S. López ◽  
Alejandro F. Alice ◽  
Horacio Heras ◽  
Emilio A. Rivas ◽  
Carmen Sánchez-Rivas
Keyword(s):  
Bacillus Subtilis ◽  
Stationary Phase ◽  
Growth Curves ◽  
Membrane Lipid ◽  
Wild Type ◽  
Lipid Packing ◽  
Anionic Phospholipids ◽  
Large Unilamellar Vesicles ◽  
Mutant Strains ◽  
Lag Period

The importance of the content of anionic phospholipids [cardiolipin (CL) and phosphatidylglycerol (PG)] in the osmotic adaptation and in the membrane structure of Bacillus subtilis cultures was investigated. Insertion mutations in the three putative cardiolipin synthase genes (ywiE, ywnE and ywjE) were obtained. Only the ywnE mutation resulted in a complete deficiency in cardiolipin and thus corresponds to a true clsA gene. The osmotolerance of a clsA mutant was impaired: although at NaCl concentrations lower than 1·2 M the growth curves were similar to those of its wild-type control, at 1·5 M NaCl (LBN medium) the lag period increased and the maximal optical density reached was lower. The membrane of the clsA mutant strain showed an increased PG content, at both exponential and stationary phase, but no trace of CL in either LB or LBN medium. As well as the deficiency in CL synthesis, the clsA mutant showed other differences in lipid and fatty acids content compared to the wild-type, suggesting a cross-regulation in membrane lipid pathways, crucial for the maintenance of membrane functionality and integrity. The biophysical characteristics of membranes and large unilamellar vesicles from the wild-type and clsA mutant strains were studied by Laurdan's steady-state fluorescence spectroscopy. At physiological temperature, the clsA mutant showed a decreased lateral lipid packing in the protein-free vesicles and isolated membranes compared with the wild-type strain. Interestingly, the lateral lipid packing of the membranes of both the wild-type and clsA mutant strains increased when they were grown in LBN. In a conditional IPTG-controlled pgsA mutant, unable to synthesize PG and CL in the absence of IPTG, the osmoresistance of the cultures correlated with their content of anionic phospholipids. The transcriptional activity of the clsA and pgsA genes was similar and increased twofold upon entry to stationary phase or under osmotic upshift. Overall, these results support the involvement of the anionic phospholipids in the growth of B. subtilis in media containing elevated NaCl concentrations.

scholarly journals The Lipid-Modifying Multiple Peptide Resistance Factor Is an Oligomer Consisting of Distinct Interacting Synthase and Flippase Subunits

mBio ◽  
2015 ◽  
Vol 6 (1) ◽  
Author(s):  
Christoph M. Ernst ◽  
Sebastian Kuhn ◽  
Christoph J. Slavetinsky ◽  
Bernhard Krismer ◽  
Simon Heilbronner ◽  
...  
Keyword(s):  
Amino Acids ◽  
Antimicrobial Agents ◽  
Membrane Surface ◽  
Integral Membrane Protein ◽  
Membrane Lipid ◽  
Host Cells ◽  
Content Type ◽  
Anionic Phospholipids ◽  
Outer Leaflet ◽  
Membrane Lipid Bilayer

ABSTRACT  Phospholipids are synthesized at the inner leaflet of the bacterial cytoplasmic membrane but have to be translocated to the outer leaflet to maintain membrane lipid bilayer composition and structure. Even though phospholipid flippases have been proposed to exist in bacteria, only one such protein, MprF, has been described. MprF is a large integral membrane protein found in several prokaryotic phyla, whose C terminus modifies phosphatidylglycerol (PG), the most common bacterial phospholipid, with lysine or alanine to modulate the membrane surface charge and, as a consequence, confer resistance to cationic antimicrobial agents such as daptomycin. In addition, MprF is a flippase for the resulting lipids, Lys-PG or Ala-PG. Here we demonstrate that the flippase activity resides in the N-terminal 6 to 8 transmembrane segments of the Staphylococcus aureus MprF and that several conserved, charged amino acids and a proline residue are crucial for flippase function. MprF protects S. aureus against the membrane-active antibiotic daptomycin only when both domains are present, but the two parts do not need to be covalently linked and can function in trans. The Lys-PG synthase and flippase domains were each found to homo-oligomerize and also to interact with each other, which illustrates how the two functional domains may act together. Moreover, full-length MprF proteins formed oligomers, indicating that MprF functions as a dimer or larger oligomer. Together our data reveal how bacterial phospholipid flippases may function in the context of lipid biosynthetic processes. IMPORTANCE  Bacterial cytoplasmic membranes are crucial for maintaining and protecting cellular integrity. For instance, they have to cope with membrane-damaging agents such as cationic antimicrobial peptides (CAMPs) produced by competing bacteria (bacteriocins), secreted by eukaryotic host cells (defensins), or used as antimicrobial therapy (daptomycin). The MprF protein is found in many Gram-positive, Gram-negative, and even archaeal commensals or pathogens and confers resistance to CAMPs by modifying anionic phospholipids with amino acids, thereby compromising the membrane interaction of CAMPs. Here we describe how MprF does not only modify phospholipids but uses an additional, distinct domain for translocating the resulting lysinylated phospholipids to the outer leaflet of the membrane. We reveal critical details for the structure and function of MprF, the first dedicated prokaryotic phospholipid flippase, which may pave the way for targeting MprF with new antimicrobials that would not kill bacteria but sensitize them to antibiotics and innate host defense molecules.

scholarly journals The Puzzling Problem of Cardiolipin Membrane-Cytochrome c Interactions: A Combined Infrared and Fluorescence Study

2021 ◽  
Vol 22 (3) ◽  
pp. 1334
Author(s):  
Francesca Ripanti ◽  
Almerinda Di Venere ◽  
Mariangela Cestelli Guidi ◽  
Martina Romani ◽  
Alessandra Filabozzi ◽  
...  
Keyword(s):  
Cytochrome C ◽  
Conformational Changes ◽  
Membrane Surface ◽  
Long Distance ◽  
Synthetic Membranes ◽  
Large Unilamellar Vesicles ◽  
Specific Order ◽  
Cyt C ◽  
Acyl Chains ◽  
Multi Phase

The interaction of cytochrome c (cyt c) with natural and synthetic membranes is known to be a complex phenomenon, involving both protein and lipid conformational changes. In this paper, we combined infrared and fluorescence spectroscopy to study the structural transformation occurring to the lipid network of cardiolipin-containing large unilamellar vesicles (LUVs). The data, collected at increasing protein/lipid ratio, demonstrate the existence of a multi-phase process, which is characterized by: (i) the interaction of cyt c with the lipid polar heads; (ii) the lipid anchorage of the protein on the membrane surface; and (iii) a long-distance order/disorder transition of the cardiolipin acyl chains. Such effects have been quantitatively interpreted introducing specific order parameters and discussed in the frame of the models on cyt c activity reported in literature.

Role of spectrin in membrane fusion and in shape change of human erythrocyte ghost

1978 ◽  
Vol 36 (2) ◽  
pp. 328-329
Author(s):  
Hideo Hayashi ◽  
Yoshikazu Hirai ◽  
John T. Penniston
Keyword(s):  
Conformational Changes ◽  
Human Erythrocyte ◽  
Shape Change ◽  
Membrane Surface ◽  
Slight Modification ◽  
Active Role ◽  
Main Role ◽  
Bank Blood ◽  
Human Erythrocyte Ghost

Spectrin is a membrane associated protein most of which properties have been tentatively elucidated. A main role of the protein has been assumed to give a supporting structure to inside of the membrane. As reported previously, however, the isolated spectrin molecule underwent self assemble to form such as fibrous, meshwork, dispersed or aggregated arrangements depending upon the buffer suspended and was suggested to play an active role in the membrane conformational changes. In this study, the role of spectrin and actin was examined in terms of the molecular arrangements on the erythrocyte membrane surface with correlation to the functional states of the ghosts.Human erythrocyte ghosts were prepared from either freshly drawn or stocked bank blood by the method of Dodge et al with a slight modification as described before. Anti-spectrin antibody was raised against rabbit by injection of purified spectrin and partially purified.

Role of PKC isozymes in water transport in toad urinary bladder

1991 ◽  
Vol 49 ◽  
pp. 302-303
Author(s):  
A.J. Mia ◽  
L.X. Oakford ◽  
T. Yorio
Keyword(s):  
Urinary Bladder ◽  
Apical Membrane ◽  
Membrane Surface ◽  
Protein A ◽  
Cell Types ◽  
Leukemic Cells ◽  
Toad Urinary Bladder ◽  
Pkc Isozymes ◽  
Antibody Labeling

Protein kinase C (PKC) isozymes, when activated, are translocated to particulate membrane fractions for transport to the apical membrane surface in a variety of cell types. Evidence of PKC translocation was demonstrated in human megakaryoblastic leukemic cells, and in cardiac myocytes and fibroblasts, using FTTC immunofluorescent antibody labeling techniques. Recently, we reported immunogold localizations of PKC subtypes I and II in toad urinary bladder epithelia, following 60 min stimulation with Mezerein (MZ), a PKC activator, or antidiuretic hormone (ADH). Localization of isozyme subtypes I and n was carried out in separate grids using specific monoclonal antibodies with subsequent labeling with 20nm protein A-gold probes. Each PKC subtype was found to be distributed singularly and in discrete isolated patches in the cytosol as well as in the apical membrane domains. To determine if the PKC isozymes co-localized within the cell, a double immunogold labeling technique using single grids was utilized.

SEM/FESEM imaging of lamellar structures in melt extruded polyethylene films

1992 ◽  
Vol 50 (2) ◽  
pp. 1142-1143
Author(s):  
R.T. Chen ◽  
M.G. Jamieson ◽  
R. Callahan
Keyword(s):  
Imaging Techniques ◽  
Membrane Surface ◽  
High Stress ◽  
Extrusion Direction ◽  
Polymeric Membranes ◽  
Scanning Tunneling ◽  
Tunneling Microscopy ◽  
Crystalline Polymers ◽  
Lamellar Structures ◽  
Resolution Imaging

“Row lamellar” structures have previously been observed when highly crystalline polymers are melt-extruded and recrystallized under high stress. With annealing to perfect the stacked lamellar superstructure and subsequent stretching in the machine (extrusion) direction, slit-like micropores form between the stacked lamellae. This process has been adopted to produce polymeric membranes on a commercial scale with controlled microporous structures. In order to produce the desired pore morphology, row lamellar structures must be established in the membrane precursors, i.e., as-extruded and annealed polymer films or hollow fibers. Due to the lack of pronounced surface topography, the lamellar structures have typically been investigated by replica-TEM, an indirect and time consuming procedure. Recently, with the availability of high resolution imaging techniques such as scanning tunneling microscopy (STM) and field emission scanning electron microscopy (FESEM), the microporous structures on the membrane surface as well as lamellar structures in the precursors can be directly examined.The materials investigated are Celgard® polyethylene (PE) flat sheet membranes and their film precursors, both as-extruded and annealed, made at different extrusion rates (E.R.).

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