Electrostatics of a peptide at a membrane/water interface. The pH dependence of melittin association with lipid vesicles

Stankowski, Stefan; Schwarz, Gerhard

doi:10.1016/0005-2736(90)90094-5

Cited by 85 publications

(45 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Such a behavior is known from a variety of cytolysins and was proposed to be based on a common cationic site that determines the lytic activity by enhancing the interaction with negatively charged vesicles or cell surfaces (17). Examples ofthese kinds ofpeptides are sarcotoxin IA, an antibacterial peptide of flesh flies (18), and melittin, the membranolytic polypeptide of the venoms from bee species (19,20).…”

Section: Discussionmentioning

confidence: 99%

Pore-forming peptide of pathogenic Entamoeba histolytica.

Leippe

Ebel

Schoenberger

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

192

133

View full text Add to dashboard Cite

A polypeptide that causes pore formation in target-cell membranes is implicated in the potent cytolytic activity of pathogenic Entamoeba histolytica. Pore-forming material was purified to apparent homogeneity by a multistep procedure, and its analysis by NaDodSO4/PAGE revealed one peptide of 4-5 kDa under nonreducing or under reducing conditions. Pore-forming activity was measured by depolarization of liposome membrane potential and was found to be optimally expressed at low pH. Active material preferentially inserted into negatively charged lipid vesicles. Treatment of purified amoeba peptide in solution or bound to liposomes with glutaraldehyde revealed oligomers upon NaDodSO4/PAGE, suggesting functionally relevant peptide-peptide interactions. The NH2-terminal amino acid sequence of the amoeba peptide was determined by protein sequencing and revealed a structural similarity to melittin, the membranolytic peptide of bee venom.

show abstract

Section: Discussionmentioning

confidence: 99%

Pore-forming peptide of pathogenic Entamoeba histolytica.

Leippe

Ebel

Schoenberger

et al. 1991

Proc. Natl. Acad. Sci. U.S.A.

192

133

View full text Add to dashboard Cite

show abstract

“…B: Digestion of NBDpost-M2 as described in Figure 6A. face active peptides Stankowski & Schwarz, 1990;Rapaport & Shai, 1991;Thiaudiere et al, 1991;Pouny et al, 1992).…”

Section: Binding Experimentsmentioning

confidence: 99%

Secondary structure, membrane localization, and coassembly within phospholipid membranes of synthetic segments derived from the N‐ and C‐termini regions of the ROMK1 K⁺ channel

Ben-Efraim¹,

Shai²

1996

Protein Science

View full text Add to dashboard Cite

The hydropathy plot of the inwardly rectifying ROMKI K+ channel, which reveals two transmembrane and a pore region domains, also reveals areas of intermediate hydrophobicity in the N terminus (MO) and in the C terminus (post"2). Peptides that correspond to MO, post"2, and a control peptide, pre-MO, were synthesized and characterized for their structure, affinity to phospholipid membranes, organizational state in membranes, and ability to self-assemble and coassemble in the membrane-bound state. CD spectroscopy revealed that both MO and post-M2 adopt highly a-helical structures in 1% SDS and 40% TWwater, whereas pre-MO is not a-helical in either 1% SDS or 40% TFEYwater. Binding experiments with NBD-labeled peptides demonstrated that both MO and post-M2, but not pre-MO, bind to zwitterionic phospholipid membranes with partition coefficients of 103-105 M". A surface localization for both post-M2 and MO was indicated by NBD shift, tryptophan quenching experiments with brominated phospholipids, and enzymatic cleavage. Resonance energy transfer measurements between fluorescently labeled pairs of donor (NBD)/ acceptor (rhodamine) peptides revealed that MO and post-M2 can coassemble in their membrane-bound state, but cannot self-associate when membrane-bound. The results are in agreement with recent data indicating that amino acids in the carboxy terminus of inwardly rectifying K+ channels have a major role in specifying the pore properties of the channels (Taglialatela M, Wible BA, Caporaso R, Brown AM, 1994, Science 264:844847; Pessia M, Bond CT, Kavanaugh MP, Adelman JP, 1995, Neuron 14:1039-1045). The relevance of the results presented herein to the suggested model for the structure of the ROMKI channel and to general aspects of molecular recognition between membrane-bound polypeptides are also discussed.Keywords: channel structure; fluorescence; K+ channels; synthetic peptides Inwardly rectifying K+ channels conduct an inward K+ current at hyperpolarizing membrane potentials. These channels, because of their rectification properties, play an important role in regulating Reprint requests to: Yechiel Shai, Department of Membrane Research and Biophysics, Weizmann Institute of Science, Rehovot, 76100 Israel; e-mail: bmshai@weizmann.weizmann.ac.il.Abbreviufions: BOC, butyloxycarbonyl; 6,7-BrPC, l-palmitoyl-2-stearoyl(6-7) dibromo-sn-glycero-3 phosphocholine; 9,IO-BrPC. l-palmitoyl-2-stearoyl(9-IO) dibromo-sn-glycero-3 phosphocholine; 11,12-BrPC, l-palmitoyl-2-stearoyl( 1 1-12) dibromo-sn-glycero-3 phosphocholine; DCC, dicyclohexylcarbodiimide; DIEA, diisopropylethylamine; DMF, dimethyl formamide; HF, hydrogen fluoride; HOBT, 1-hydroxybenzotriazole; LUV, large unilamellar vesicles; NBD, 7-nitrobenz-2-oxa-l,3-diazole, 4-yl; Pam, (pheny1acetamido)methyl; NBD-F, 4-fluoro-7-nitrobenz-2-oxa-1,3-diazole; PC, egg phosphatidylcholine; Rho, tetramethylrhodamine; Rho-Su, 5-(and-6)-carboxytetramethylrhodamine succinimidyl ester; RP-HPLC, reversephase HPLC; SUV, small unilamellar vesicles; TFA, trifluoroacetic acid; TFE, tri...

show abstract

“…3. For DNCmelittin the fitted line turns out to be: The partition coefficient is given in Table 2 , being slightly lower than those reported in the literature [10,12,15,19,22,31,42]. An effective charge smaller than that expected from the number of ionizable groups is a well-known phenomenon, previously described for melittin (+6 e.u.)…”

Section: Binding Of Dnc-melittin To Neutral Bilayersmentioning

confidence: 64%

“…+1.5 e.u. per melittin molecule on the surface [12], the formation of aggregates seems to be reasonable at higher peptide concentrations. From the obtained data, it is not possible to distinguish whether melittin binds as monomer to the surface forming multilayers at higher concentrations, or whether melittin tetramers, which are already formed in solution at higher concentrations, adsorb to the surface as tetramers building multilayers.…”

Section: As a First Approximation A Value Ofmentioning

confidence: 82%