Binding of Peptides with Basic and Aromatic Residues to Bilayer Membranes

Zhang, Wenyi; Crocker, Evan; McLaughlin, Stuart; Smith, Steven O.

doi:10.1074/jbc.m301652200

Cited by 109 publications

(62 citation statements)

References 55 publications

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“…Clusters of basic and aromatic residues in general are found in a wide range of membrane-associated proteins involved in signal transduction, and Phe residues are often associated with basic residues in peripheral membrane proteins (42). Compared with Leu, Phe has ϳ12% larger hydrophobic surface and, as opposed to Trp and Tyr, has no preference for the interface over the core of the membrane.…”

Section: Discussionmentioning

confidence: 99%

Temporin-SHf, a New Type of Phe-rich and Hydrophobic Ultrashort Antimicrobial Peptide

Abbassi¹,

Lequin

Piesse

et al. 2010

Journal of Biological Chemistry

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Because issues of cost and bioavailability have hampered the development of gene-encoded antimicrobial peptides to combat infectious diseases, short linear peptides with high microbial cell selectivity have been recently considered as antibiotic substitutes. A new type of short antimicrobial peptide, designated temporin-SHf, was isolated and cloned from the skin of the frog Pelophylax saharica. Temporin-SHf has a highly hydrophobic sequence (FFFLSRIFa) and possesses the highest percentage of Phe residues of any known peptide or protein. Moreover, it is the smallest natural linear antimicrobial peptide found to date, with only eight residues. Despite its small size and hydrophobicity, temporin-SHf has broad-spectrum microbicidal activity against Gram-positive and Gram-negative bacteria and yeasts, with no hemolytic activity. CD and NMR spectroscopy combined with restrained molecular dynamics calculations showed that the peptide adopts a well defined non-amphipathic ␣-helical structure from residue 3 to 8, when bound to zwitterionic dodecyl phosphocholine or anionic SDS micelles. Relaxation enhancement caused by paramagnetic probes showed that the peptide adopts nearly parallel orientations to the micelle surface and that the helical structure is stabilized by a compact hydrophobic core on one face that penetrates into the micelle interior. Differential scanning calorimetry on multilamellar vesicles combined with membrane permeabilization assays on bacterial cells indicated that temporin-SHf disrupts the acyl chain packing of anionic lipid bilayers, thereby triggering local cracks and microbial membrane disintegration through a detergent-like effect probably via the carpet mechanism. The short length, compositional simplicity, and broad-spectrum activity of temporin-SHf make it an attractive candidate to develop new antibiotic agents.

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Section: Discussionmentioning

confidence: 99%

Temporin-SHf, a New Type of Phe-rich and Hydrophobic Ultrashort Antimicrobial Peptide

Abbassi¹,

Lequin

Piesse

et al. 2010

Journal of Biological Chemistry

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“…Arginines and lysines within the BHC and anionic phospholipid headgroups are additionally required for membrane binding. This synergism of charged and hydrophobic components is typical of the membrane binding mechanisms of other basic/ hydrophobic clusters (29,(31)(32)(33)(34), with the electrostatic force between the basic amino acid side chains and the anionic head-FIGURE 6. PT domain co-sediments with phospholipid vesicles containing PI(3,4,5)P 3 .…”

Section: Discussionmentioning

confidence: 99%

“…Mutation of Phe 723 had no noticeable effect on its own (RG1-F in Fig. 1, A and D) Bilayer penetration of aromatic side chains is often coupled with electrostatic interactions between neighboring basic side chains and phospholipid headgroups (34). There is a conserved cluster of basic residues in the N-terminal portion of the PT domain as follows: three arginines or lysines at positions 719 -721 and a lysine at 725, adjacent to the tryptophan (Fig.…”

Section: Basic/hydrophobic Cluster In the Pt Domain Mediatesmentioning

confidence: 99%

Phosphoinositide 3-Kinase Regulates Plasma Membrane Targeting of the Ras-specific Exchange Factor RasGRP1

Zahedi

Goo

Beaulieu

et al. 2011

Journal of Biological Chemistry

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Receptor-induced targeting of exchange factors to specific cellular membranes is the predominant mechanism for initiating and compartmentalizing signal transduction by Ras GTPases. The exchange factor RasGRP1 has a C1 domain that binds the lipid diacylglycerol and thus can potentially mediate membrane localization in response to receptors that are coupled to diacylglycerol-generating phospholipase Cs. However, the C1 domain is insufficient for targeting RasGRP1 to the plasma membrane. We found that a basic/hydrophobic cluster of amino acids within the plasma membrane-targeting domain of RasGRP1 is instead responsible for plasma membrane targeting. This basic/hydrophobic cluster binds directly to phospholipid vesicles containing phosphoinositides via electrostatic interactions with polyanionic phosphoinositide headgroups and insertion of a tryptophan into the lipid bilayer. B cell antigen receptor ligation and other stimuli induce plasma membrane targeting of RasGRP1 by activating the phosphoinositide 3-kinase signaling pathway, which generates phosphoinositides within the plasma membrane. Direct detection of phosphoinositides by the basic/ hydrophobic cluster of RasGRP1 provides a novel mechanism for coupling and co-compartmentalizing phosphoinositide 3-kinase and Ras signaling and, in coordination with diacylglycerol detection by the C1 domain, gives RasGRP1 the potential to serve as an integrator of converging signals from the phosphoinositide 3-kinase and phospholipase C pathways.Ras GTPases are membrane-bound signal-transducing proteins that control a wide range of cellular responses to external stimuli. Their attachment to specific cellular membranes allows Ras GTPases to confine, concentrate, and organize networks of signal detectors and transmitters (1). Guanine nucleotide exchange factors directly activate Ras GTPases by promoting their conversion to the active GTP-bound state (2). Additionally, exchange factors regulate Ras activation by detecting and integrating inputs from signal transducers coupled to cell-surface receptors. Conversion of receptor-initiated signals into Ras activation is often achieved by spatial regulation of an exchange factor (2, 3). In the absence of signal detection, the exchange factor is sequestered away from its Ras GTPase substrates, whereas the activating signal induces membrane binding of the exchange factor and thus co-localizes it with Ras GTPase substrates. RasGRP1 is a Ras-activating exchange factor that is expressed in many cell types and can be activated by diverse receptors (2, 4). In lymphocytes, initiation of Ras signaling by RasGRP1 controls progression through developmental and immunoselection checkpoints, whereas misregulation of RasGRP1 can trigger autoimmunity or oncogenic transformation (5-10). Depending on the quality and intensity of signaling from receptors, RasGRP1 can be specifically targeted to the plasma membrane (11)(12)(13)(14)(15) or to endomembranes such as Golgi or the endoplasmic reticulum (11, 13, 16 -19). The site of RasGRP1 localization can h...

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“…The former model seems more plausible, since mNG-MARCKS also shows preference for Ld, although this preference is less extreme. The MARCKS peptide comprises not only 13 basic residues, which presumably are unstructured, but also 5 phenylalanine residues that might insert into the membrane (63,85,92). Mutational studies could address the mechanism of Ld preference for these two proteins.…”

Section: Discussionmentioning

confidence: 99%

Effects of Membrane Charge and Order on Membrane Binding of the Retroviral Structural Protein Gag

Wen

Dick

Feigenson

et al. 2016

J Virol

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The retroviral structural protein Gag binds to the inner leaflet of the plasma membrane (PM), and many cellular proteins do so as well. We used Rous sarcoma virus (RSV) Gag together with membrane sensors to study the principles governing peripheral protein membrane binding, including electrostatics, specific recognition of phospholipid headgroups, sensitivity to phospholipid acyl chain compositions, preference for membrane order, and protein multimerization. We used an in vitro liposome-pelleting assay to test protein membrane binding properties of Gag, the well-characterized MARCKS peptide, a series of fluorescent electrostatic sensor proteins (mNG-KRn), and the specific phosphatidylserine (PS) binding protein Evectin2. RSV Gag and mNG-KRn bound well to membranes with saturated and unsaturated acyl chains, whereas the MARCKS peptide and Evectin2 preferentially bound to membranes with unsaturated acyl chains. To further discriminate whether the primary driving force for Gag membrane binding is electrostatic interactions or preference for membrane order, we measured protein binding to giant unilamellar vesicles (GUVs) containing the same PS concentration in both disordered (Ld) and ordered (Lo) phases. RSV Gag and mNG-KRn membrane association followed membrane charge, independent of membrane order. Consistent with pelleting data, the MARCKS peptide showed preference for the Ld domain. Surprisingly, the PS sensor Evectin2 bound to the PS-rich Ld domain with 10-fold greater affinity than to the PS-rich Lo domain. In summary, we found that RSV Gag shows no preference for membrane order, while proteins with reported membrane-penetrating domains show preference for disordered membranes. IMPORTANCERetroviral particles assemble on the PM and bud from infected cells. Our understanding of how Gag interacts with the PM and how different membrane properties contribute to overall Gag assembly is incomplete. This study examined how membrane charge and membrane order influence Gag membrane association. Consistent with previous work on RSV Gag, we report here that electrostatic interactions provide the primary driving force for RSV Gag membrane association. Using phase-separated GUVs with known lipid composition of the Ld and Lo phases, we demonstrate for the first time that RSV Gag is sensitive to membrane charge but not membrane order. In contrast, the cellular protein domain MARCKS and the PS sensor Evectin2 show preference for disordered membranes. We also demonstrate how to define GUV phase composition, which could serve as a tool in future studies of protein membrane interactions.T he retroviral structural protein Gag provides the primary driving force for virus assembly at the inner leaflet of the plasma membrane (PM), and Gag alone is sufficient to assemble into virus-like particles (VLPs) in cells (1) and also in vitro (2). Gag is synthesized in the cytosol and then is targeted to the PM by the N-terminal domain, MA. How Gag interacts with the inner leaflet of the PM and how the biophysical properties of the PM...

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Binding of Peptides with Basic and Aromatic Residues to Bilayer Membranes

Cited by 109 publications

References 55 publications

Temporin-SHf, a New Type of Phe-rich and Hydrophobic Ultrashort Antimicrobial Peptide

Temporin-SHf, a New Type of Phe-rich and Hydrophobic Ultrashort Antimicrobial Peptide

Phosphoinositide 3-Kinase Regulates Plasma Membrane Targeting of the Ras-specific Exchange Factor RasGRP1

Effects of Membrane Charge and Order on Membrane Binding of the Retroviral Structural Protein Gag

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