emerimicins and antiamoebin

Amoebiasis is a parasitic infectious disease caused by the enteric protozoan Entamoeba histolytica, a leading basis of deaths accounted to parasites, succeeding malaria and schistosomiasis. Conventional treatment methodologies used to deal with amoebiasis mainly rely on the administration of anti-amoebic compounds and vaccines but are often linked with substantial side-effects on the patient. Besides, cases of development of drug resistance in protozoans have been recorded, contributing further to the reduction in the efficiency of the treatment. Loopholes in the efficacious management of the disease call for the development of novel methodologies to manage amoebiasis. A way to achieve this is by targeting the essential metabolic processes of 'encystation' and 'excystation', and the associated biomolecules, thus interrupting the biphasic life cycle of the parasite. Technologies like the CRISPR-Cas9 system can efficiently be exploited to discover novel and essential molecules that regulate the protozoan's metabolism, while efficiently manipulating and managing the known drug targets, leading to an effective halt and forestall to the enteric infection. This review presents a perspective on these essential metabolic processes and the associated molecules that can be targeted efficaciously to prevent the transmission of amoebiasis, thus managing the disease and proving to be a fruitful endeavour.

Topics: Amebiasis; Animals; Chitinases; Entamoeba histolytica; Entamoebiasis; Humans; Lectins; Models, Biological; Molecular Conformation; Molecular Targeted Therapy; Peptaibols; Signal Transduction

Kinks or bends introduced in peptides and proteins by "helical distorter" residues such as proline, other imino acids and glycine, especially when these are in close proximity in the sequence, are increasingly recognized as playing an essential role in the gating of channel-forming peptides as well as of physiological ion channels. Peptaibols are useful simple models for the much more complex biological ion channels, especially voltage-gated ones. In this short review, we compare the monomeric structures of three selected peptaibols (alamethicin, trichotoxin and antiamoebin) that widely differ with regards their near-central kink angles and dipolar moment orientations. These structural features are then shown to be correlated to the different patterns of channel activity, both at the macroscopic and single-channel levels of investigation.

Topics: Alamethicin; Antimicrobial Cationic Peptides; Electrophysiology; Ion Channels; Kinetics; Lipid Bilayers; Models, Molecular; Peptaibols; Peptides; Protein Structure, Secondary

Antiamoebin I (Aam-I) is a membrane-active peptaibol antibiotic isolated from fungal species belonging to the genera Cephalosporium, Emericellopsis, Gliocladium, and Stilbella. In comparison with other 16-amino acid-residue peptaibols, e.g., zervamicin IIB (Zrv-IIB), Aam-I possesses relatively weak biological and channel-forming activities. In MeOH solution, Aam-I demonstrates fast cooperative transitions between right-handed and left-handed helical conformation of the N-terminal (1-8) region. We studied Aam-I spatial structure and backbone dynamics in the membrane-mimicking environment (DMPC/DHPC bicelles)(1) ) by heteronuclear (1) H,(13) C,(15) N-NMR spectroscopy. Interaction with the bicelles stabilizes the Aam-I right-handed helical conformation retaining significant intramolecular mobility on the ms-μs time scale. Extensive ms-μs dynamics were also detected in the DPC and DHPC micelles and DOPG nanodiscs. In contrast, Zrv-IIB in the DPC micelles demonstrates appreciably lesser mobility on the μs-ms time scale. Titration with Mn(2+) and 16-doxylstearate paramagnetic probes revealed Aam-I binding to the bicelle surface with the N-terminus slightly immersed into hydrocarbon region. Fluctuations of the Aam-I helix between surface-bound and transmembrane (TM) state were observed in the nanodisc membranes formed from the short-chain (diC12 : 0) DLPC/DLPG lipids. All the obtained experimental data are in agreement with the barrel-stave model of TM pore formation, similarly to the mechanism proposed for Zrv-IIB and other peptaibols. The observed extensive intramolecular dynamics explains the relatively low activity of Aam-I.

Topics: Lipids; Magnetic Resonance Spectroscopy; Micelles; Models, Molecular; Nanostructures; Peptaibols; Peptides; Porosity; Protein Structure, Secondary; Solubility; Water

The two segments, 1-9 and 10-16, of the peptaibol antibiotic antiamoebin I, i.e., the nonapeptide Ac-Phe-Aib-Aib-Aib-D,L-Iva-Gly-Leu-Aib-Aib-OH (15) and the heptapeptide Z-Hyp-Gln-D,L-Iva-Hyp-Aib-Pro-Pheol (34), have been prepared as mixtures of the epimers containing D,L-Iva. All α,α-disubstituted α-amino acids were introduced by the 'azirine/oxazolone method', in which amino or peptide acids are coupled with the corresponding 2H-azirin-3-amines, followed by selective hydrolysis of the terminal amide bond. The amino acids Hyp and Gln were introduced as Z-protected(4) ) (2S,4R)-4-(tert-butoxy)proline (19) and methyl N-[bis(4-methoxyphenyl)methyl]glutamine (26). Coupling of peptide segments was achieved via the 'mixed anhydride' method, the DCC/HOBt or TBTU/HOBt strategy. The crystal structure of the segment 6-9 was determined by X-ray crystallography and displayed the presence of a β-turn conformation.

Topics: Amino Acid Sequence; Azirines; Combinatorial Chemistry Techniques; Crystallography, X-Ray; Molecular Sequence Data; Oxazolone; Peptaibols; Peptides

Distinctions between isobaric residues have been a major challenge in mass spectrometric peptide sequencing. Here, we propose a methodology for distinction among isobaric leucine, isoleucine, and hydroxyproline, a commonly found post-translationally modified amino acid with a nominal mass of 113 Da, through a combined electron transfer dissociation-collision-induced dissociation approach. While the absence of c and z(•) ions, corresponding to the Yyy-Xxx (Xxx = Leu, Ile, or Hyp) segment, is indicative of the presence of hydroxyproline, loss of isopropyl (Δm = 43 Da) or ethyl radicals (Δm = 29 Da), through collisional activation of z radical ions, are characteristic of leucine or isoleucine, respectively. Radical migration processes permit distinctions even in cases where the specific z(•) ions, corresponding to the Yyy-Leu or -Ile segments, are absent or of low intensity. This tandem mass spectrometric (MS(n)) method has been successfully implemented in a liquid chromatography-MS(n) platform to determine the identity of 23 different isobaric residues from a mixture of five different peptides. The approach is convenient for distinction of isobaric residues from any crude peptide mixture, typically encountered in natural peptide libraries or proteomic analysis.

Topics: Amino Acid Sequence; Biological Products; Hydroxyproline; Isoleucine; Leucine; Mass Spectrometry; Melitten; Molecular Sequence Data; Peptaibols; Peptides; Sequence Analysis, Protein; Wasp Venoms

Molecular-dynamics simulations were carried out to ascertain which of the potential multimeric forms of the transmembrane peptaibol channel, antiamoebin, is consistent with its measured conductance. Estimates of the conductance obtained through counting ions that cross the channel and by solving the Nernst-Planck equation yield consistent results, indicating that the motion of ions inside the channel can be satisfactorily described as diffusive. The calculated conductance of octameric channels is markedly higher than the conductance measured in single channel recordings, whereas the tetramer appears to be nonconducting. The conductance of the hexamer was estimated to be 115 ± 34 pS and 74 ± 20 pS, at 150 mV and 75 mV, respectively, in satisfactory agreement with the value of 90 pS measured at 75 mV. On this basis, we propose that the antiamoebin channel consists of six monomers. Its pore is large enough to accommodate K⁺ and Cl⁻ with their first solvation shells intact. The free energy barrier encountered by K⁺ is only 2.2 kcal/mol whereas Cl⁻ encounters a substantially higher barrier of nearly 5 kcal/mol. This difference makes the channel selective for cations. Ion crossing events are shown to be uncorrelated and follow Poisson statistics.

Topics: Biomechanical Phenomena; Diffusion; Electric Conductivity; Ion Channel Gating; Ion Channels; Ion Transport; Lipids; Molecular Dynamics Simulation; Peptaibols; Peptides; Pliability; Porosity; Protein Stability; Protein Structure, Quaternary; Structure-Activity Relationship; Thermodynamics; Water

High-resolution NMR is shown to be applicable for investigation of membrane proteins and membrane-active peptides embedded into lipid-protein nanodiscs (LPNs). (15)N-Labeled K+-channel from Streptomyces lividans (KcsA) and the antibiotic antiamoebin I from Emericellopsis minima (Aam-I) were embedded in LPNs of different lipid composition. Formation of stable complexes undergoing isotropic motion in solution was confirmed by size-exclusion chromatography and (31)P-NMR spectroscopy. The 2D 1H-(15)N-correlation spectra were recorded for KcsA in the complex with LPN containing DMPC and for Aam-I in LPNs based on DOPG, DLPC, DMPC, and POPC. The spectra recorded were compared with those in detergent-containing micelles and small bicelles commonly used in high-resolution NMR spectroscopy of membrane proteins. The spectra recorded in LPN environments demonstrated similar signal dispersion but significantly increased (1)H(N) line width. The spectra of Aam-I embedded in LPNs containing phosphatidylcholine showed significant selective line broadening, thus suggesting exchange process(es) between several membrane-bound states of the peptide. (15)N relaxation rates were measured to obtain the effective rotational correlation time of the Aam-I molecule. The obtained value (approximately 40 nsec at 45 degrees C) is indicative of additional peptide motions within the Aam-I/LPN complex.

Topics: Bacterial Proteins; Hypocreales; Lipids; Magnetic Resonance Spectroscopy; Membrane Proteins; Nanostructures; Peptaibols; Peptides; Potassium Channels

Topics: Amino Acid Sequence; Apolipoprotein A-I; Lipid Bilayers; Membrane Proteins; Molecular Sequence Data; Nanostructures; Nuclear Magnetic Resonance, Biomolecular; Peptaibols; Peptides; Phosphatidylcholines; Phosphatidylglycerols; Phospholipids

Antiamoebin I (Aam-I) is a membrane-active peptaibol antibiotic isolated from fungal species belonging to the genera Cephalosporium, Emericellopsis, Gliocladium, and Stilbella. Antiamoebin I has the amino acid sequence: Ac-Phe(1)-Aib-Aib-Aib-Iva-Gly-Leu-Aib(8)-Aib-Hyp-Gln-Iva-Hyp-Aib-Pro-Phl(16). By using the uniformly (13)C,(15)N-labeled sample of Aam-I, the set of conformationally dependent J couplings and (3h)J(NC) couplings through H-bonds were measured. Analysis of these data along with the data on magnetic nonequivalence of the (13)C(beta) nuclei (Deltadelta((13)C(beta))) in Aib and Iva residues allowed us to draw the univocal conclusion that the N-terminal part (Phe(1)-Gly(6)) of Aam-I in MeOH solution is in fast exchange between the right-handed and left-handed 3(10)-helical conformations, with an approximately equal population of both states. An additional conformational exchange process was found at the Aib(8) residue. The (15)N-NMR-relaxation and CD-spectroscopy measurements confirmed these findings. Molecular modeling and Monte Carlo simulations revealed that both exchange processes are correlated and coupled with significant hinge-bending motions around the Aib(8) residue. Our results explain relatively low activity of Aam-I with respect to other 15-amino acid residue peptaibols (for example, zervamicin) in functional and biological tests. The high dynamic 'propensity' possibly prevents both initial binding of the antiamoebin to the membrane and subsequent formation of stable ionic channels according to the barrel-stave mechanism.

Topics: Circular Dichroism; Methanol; Monte Carlo Method; Nuclear Magnetic Resonance, Biomolecular; Peptaibols; Peptides; Protein Conformation; Solutions; Stereoisomerism

Antiamoebins I, III and XVI as well as several others in minor amounts were produced by four strains of the coprophilous fungus Stilbella erythrocephala (syn. S. fimetaria) in its natural substrate and in liquid culture. The total antiamoebin concentration in dung was 126-624 microg g(-1) fresh weight, with minimum inhibitory concentrations against most other coprophilous fungi being at or below 100 microg mL(-1). Myrocin B, not previously described from S. erythrocephala, was also produced, but only at low, nonfungicidal levels (< 5.3 microg g(-1)). No other antifungal substances were detected. It is concluded that antiamoebins are responsible for antibiosis in dung colonized by S. erythrocephala.

Topics: Antibiosis; Diterpenes; Fungi; Manure; Peptaibols; Peptides

Permeabilization of the phospholipid membrane, induced by the antibiotic peptides zervamicin IIB (ZER), ampullosporin A (AMP) and antiamoebin I (ANT) was investigated in a vesicular model system. Membrane-perturbing properties of these 15/16 residue peptides were examined by measuring the K(+) transport across phosphatidyl choline (PC) membrane and by dissipation of the transmembrane potential. The membrane activities are found to decrease in the order ZER>AMP>>ANT, which correlates with the sequence of their binding affinities. To follow the insertion of the N-terminal Trp residue of ZER and AMP, the environmental sensitivity of its fluorescence was explored as well as the fluorescence quenching by water-soluble (iodide) and membrane-bound (5- and 16-doxyl stearic acids) quenchers. In contrast to AMP, the binding affinity of ZER as well as the depth of its Trp penetration is strongly influenced by the thickness of the membrane (diC(16:1)PC, diC(18:1)PC, C(16:0)/C(18:1)PC, diC(20:1)PC). In thin membranes, ZER shows a higher tendency to transmembrane alignment. In thick membranes, the in-plane surface association of these peptaibols results in a deeper insertion of the Trp residue of AMP which is in agreement with model calculations on the localization of both peptide molecules at the hydrophilic-hydrophobic interface. The observed differences between the membrane affinities/activities of the studied peptaibols are discussed in relation to their hydrophobic and amphipathic properties.

Topics: Anti-Bacterial Agents; Hydrophobic and Hydrophilic Interactions; Ion Channels; Lipid Bilayers; Membrane Potentials; Peptaibols; Peptides; Permeability; Spectrometry, Fluorescence

Antiamoebin I is a membrane-active peptaibol produced by fungi of the species Emericellopsis which is capable of forming ion channels in membranes. Previous structure determinations by x-ray crystallography have shown the molecule is mostly helical, with a deep bend in the center of the polypeptide, and that the backbone structure is independent of the solvent used for crystallization. In this study, the solution structure of antiamoebin was determined by NMR spectroscopy in methanol, a solvent from which one of the crystal structures was determined. The ensemble of structures produced exhibit a right-handed helical C terminus and a left-handed helical conformation toward the N-terminus, in contrast to the completely right-handed helices found in the crystal structures. The NMR results also suggest that a "hinge" region exists, which gives flexibility to the polypeptide in the central region, and which could have functional implications for the membrane insertion process. A model for the membrane insertion and assembly process is proposed based on the antiamoebin solution and crystal structures, and is contrasted with the assembly and insertion mechanism proposed for other ion channel-forming polypeptides.

Topics: Anti-Bacterial Agents; Computer Simulation; Crystallography; Hydrogen Bonding; Ion Channels; Models, Molecular; Nuclear Magnetic Resonance, Biomolecular; Peptaibols; Peptides; Protein Conformation; Protein Denaturation; Protein Folding; Protein Structure, Secondary; Protein Structure, Tertiary; Solutions

Malaria caused by Plasmodium falciparum is a major public health problem in the developing countries of the world. Clinical treatment of malaria has become complicated due to the occurrence of infections caused by drug resistant parasites. Secondary metabolites from fungi are an attractive source of chemotherapeutic agents. This work reports the isolation and in vitro antiplasmodial activities of peptide antibiotics of fungal origin. The three peptide antibiotics used in this study were efrapeptins, zervamicins, and antiamoebin. The high-performance liquid chromatography-purified peptides were characterized by nuclear magnetic resonance and mass spectral analysis. All three fungal peptides kill P. falciparum in culture with 50% inhibitory concentrations in the micromolar range. A possible mode of action of these peptide antibiotics on P. falciparum is presented.

Topics: Amino Acid Sequence; Animals; Anti-Bacterial Agents; Antimalarials; Ascomycota; Erythrocytes; Hypocreales; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Peptaibols; Peptides; Plasmodium falciparum; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Mixtures of the microheterogeneous 16-mer peptaibol antibiotics called antiamoebins (AAM) have been isolated from the culture broths of strains of the filamentous fungi Stilbella erythrocephala ATCC 28144, Stilbella fimetaria CBS 548.84 and Gliocladium catenulatum CBS 511.66. Sequences were determined using on-line HPLC together with positive- and negative-ion electrospray ionization mass spectrometry. Some characteristic features are recognized in the mass spectrometric fragmentation pattern of AAM. From a sample originally used for sequencing AAM (from Hindustan Antibiotics, Ltd., Pimpri, Poona-411018, India), and a sample of AAM commercially available (from Sigma Chemicals, St. Louis, MO, USA) HPLC elution profiles and sequences were assigned. Further, sequences of AAM previously isolated from Emericellopsis synnematicola CBS 176.60 and Emericellopsis salmosynnemata CBS 382.62 were determined. The peptide designated AAM I was the most abundant in all isolates and its structure could be confirmed. AAM II was detectable as a minor component (1.9%) only in the original sample of AAM, but not in the other isolates. The structures of AAM III, IV and V, which had previously been partly assigned, were definitely established, and the new sequences AAM VI-XVI were elucidated. AAM showing Phe1/Leu1 or Phe1/Val1 exchange, respectively, are produced in amounts only by S. erythrocephala. Sequences, HPLC elution profiles ('fingerprints') and relative amounts of peptides of all isolates were correlated.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Chromatography, High Pressure Liquid; Fermentation; Fungi; Mass Spectrometry; Mitosporic Fungi; Molecular Sequence Data; Molecular Structure; Peptaibols; Peptides

The successful use of molecular-replacement methods for the solution of the intermediate-sized helical polypeptide antiamoebin I required the careful consideration of a number of parameters and exhibited some unusual characteristics when compared with molecular-replacement solutions of globular proteins. High-resolution data were required owing to several features, including the comma-like shape of the molecule (which results in a pseudo-symmetric structure at low resolution), the relative uniformity of the structure in the direction along the helix axis and the small differences between the two independent molecules in the P1 asymmetric unit. Other parameters which were important for the solution of this relatively low solvent content closely packed cell included the radius of integration, the use of normalized structure factors and especially the choice of starting model.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Crystallography, X-Ray; Models, Molecular; Molecular Sequence Data; Peptaibols; Peptides; Protein Conformation; Protein Structure, Secondary; Solutions

Crystals of an ion-channel-forming peptaibol peptide in a partial membrane environment have been obtained by cocrystallizing antiamoebin with n-octanol. The antiamoebin molecule has a bent helical conformation very similar to that established for Leu-zervamicin, despite a significantly different sequence for residues 1-8. The bent helices assemble to form a polar channel in the shape of an hour glass that is quite comparable to that of Leu-zervamicin. The molecules of cocrystallized octanol are found in two different areas with respect to the assembly of peptide molecules. One octanol molecule mimics a membrane segment along the hydrophobic exterior of the channel assembly. The other octanol molecules fill the channel in such a way that their OH termini satisfy the C==O moieties directed into the interior of the channel. Structure parameters for C82 H27 N17 O20(.3) C8H18O are space group P2(1) 2(1) 2(1), a = 9.143(2) A, b = 28.590(8) A, c = 44.289(8) A, Z = 4, agreement factor R1 = 11.95% for 4,113 observed reflections [>4sigma(F)], resolution approximately 1.0 A.

Topics: Anti-Bacterial Agents; Crystallography, X-Ray; Ion Channels; Models, Molecular; Molecular Mimicry; Peptaibols; Peptides; Protein Conformation

Antiamoebin is a member of the peptaibol family of polypeptides and has a unique antibiotic activity: it acts as an antiamoebic agent, but does not effectively haemolyze erythrocytes even though it does exhibit membrane-modifying activity.. The structure of antiamoebin I has been determined by X-ray crystallography at 1.4 A resolution. The molecule forms a helical structure, which, as a result of the presence of a number of proline and hydroxyproline residues, has a deep bend in the middle. Circular dichroism spectroscopy, single-channel conductance studies and fluorescence diffusion studies suggest a mode of ion transport that is entirely different from that of the other two members of the peptaibol family (alamethicin and zervamicin) whose structures and functions have been examined in detail.. The structure of the polypeptide has been determined and a functional model for its mode of action in membranes is presented. Although under some conditions antiamoebin may form ion channels, unlike the closely related alamethicin and zervamicin polypeptides, its major membrane-modifying activity appears to be as an ion carrier.

Topics: Alamethicin; Amebicides; Anti-Bacterial Agents; Carrier Proteins; Circular Dichroism; Crystallography, X-Ray; Membranes; Models, Chemical; Models, Molecular; Peptaibols; Peptides; Proline

Antiamoebin is a 16-residue polypeptide whose crystal structure and lytic activity in membrane vesicles have recently been reported. It is a bent helical molecule and a member of the peptaibol family of antibiotics. Under conditions which produce voltage-dependent conductance activity by other members of the family, no single-channel conductance was detected for antiamoebin, and a carrier-like mechanism was put forward to account for its mode of action. We now present evidence for pore formation that is largely voltage-insensitive, with large amplitude single-channel events on top of a background conductance that may account for the previously proposed carrier-like activity. Thus, antiamoebin may be the first instance of a peptide which can function both as an ion carrier and a pore former.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Carrier Proteins; Ion Channel Gating; Lipid Bilayers; Molecular Sequence Data; Peptaibols; Peptides

The crystal structure of the nonapeptide Boc-D-Phe-Aib-Aib-Aib-Aib-Gly-Leu-Aib-Aib-OMe (I), which is an analogue of the N-terminal sequence of antiamoebins and emerimicins, establishes a completely 3(10)-helical conformation with seven successive intramolecular 4-->1 hydrogen bonds. The average, phi,psi values for residues 1-8 are -59 degrees and -32 degrees, respectively. Crystal parameters are C47H77N9O12, space group P1, a = 10.636 (4) A, b = 11.239 (4) A, c = 12.227 (6) A, alpha = 101.17 (4) degrees, beta = 97.22 (4) degrees, gamma = 89.80 (3) degrees, Z = 1, R = 5.95% for 3018 data with magnitude of F0 > 3 sigma(F), resolution 0.93 A. The use of the torsion angle kappa = C(i-1)N(i)C alpha(i)C beta(i), where kappa = 68 degrees for D-Phe and kappa = 164 degrees for L-Leu, confirms the opposite configurations of these residues. The phi,psi values of -62 degrees and -32 degrees at D-Phe are unusual, since this region is characteristic of residues with L configurations. Peptide I possesses only two chiral residues of opposing configuration. The observed right-handed 3(10)-helical structure suggests that helix sense has probably been determined by the stereochemical preferences of the Leu residue.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Hydrogen Bonding; Molecular Sequence Data; Oligopeptides; Peptaibols; Peptides; Peptides, Cyclic; Protein Conformation; Protein Structure, Secondary

The effect of different representatives of the group of peptaibiotics, alpha-amino-isobutyric acid rich secondary metabolites of filamentous fungi, on Culex pipiens larvae was studied. Light and transmission electron microscopy techniques were used to localize the intracellular damage and to determine the target organells for the mode of action of peptaibols in mosquito larvae. Though different in insecticidal activity, all tested compounds induced the same type of tissue damage, which was characterized by heavy challenge of mitochondria followed by partial swelling, crystaeolysis and destruction of mitochondrial walls. It is concluded that the mode of action of peptaibols in mosquito larvae is mediated through the damage of mitochondria. The structure-mosquitocidal effect of these compounds, their potential mode of action and role in the natural fungal entomopathogenic process are briefly discussed.

Topics: Alamethicin; Amino Acid Sequence; Aminoisobutyric Acids; Animals; Anti-Bacterial Agents; Antimicrobial Cationic Peptides; Culicidae; Fungi; Insecticides; Intercellular Signaling Peptides and Proteins; Larva; Mitochondria; Molecular Sequence Data; Organic Chemicals; Peptaibols; Peptides; Peptides, Cyclic

Trypanosomiasis is a parasitic disease, prevailing in both humans and animals, caused by a single-cell parasite, Trypanosoma spp. Three microbial metabolites, namely antiamoebin, F-857 and 6-MFA, were evaluated for trypanocidal activity by using a mouse model of trypanosomiasis, which is caused by T. evansi. The significance of the biological and biochemical parameters with respect to physio-pathology of trypanosomiasis and their implications in the evaluation of new trypanocidal compounds were discussed.

Topics: Animals; Anti-Bacterial Agents; Blood Glucose; Disease Models, Animal; Fungal Proteins; Mice; Peptaibols; Peptides; Peptides, Cyclic; Trypanocidal Agents; Trypanosomiasis

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Calcium; Cell Membrane Permeability; Lipid Bilayers; Molecular Sequence Data; Peptaibols; Peptide Fragments; Peptides; Peptides, Cyclic

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Peptaibols; Peptide Fragments; Peptides; Peptides, Cyclic; Protein Conformation

The conformations of the 16-residue fungal peptide antiamoebin I (Ac-Phe-Aib-Aib-Aib-D-Iva-Gly-Leu-Aib-Aib-Hyp-Gln-D-Iva-Hyp-Aib-Pro-P hol) have been investigated in dimethyl sulfoxide solution by one- and two-dimensional NMR techniques. A substantial number of resonances in the 270-MHz 1H NMR spectrum have been assigned. Intramolecularly hydrogen-bonded (solvent inaccessible) NH groups have been identified by determining solvent and temperature dependence of NH chemical shifts and rates of hydrogen-deuterium exchange. Ten backbone NH groups are inaccessible to solvent, while three NH groups assigned to the Phe(1), Aib(2), and Aib(8) residues are exposed to solvent. Interresidue nuclear Overhauser effects are consistent with psi values of approximately 120 +/- 30 degrees for Phe(1) and Leu(7). The NMR results, together with the stereochemical constraints imposed by the presence of alpha-aminoisobutyryl, isovalyl, prolyl, and 4-hydroxyprolyl residues, favor a highly ordered structure. Two backbone conformations consistent with the data are considered. Antiamoebin is shown to be an effective uncoupler of oxidative phosphorylation in rat liver mitochondria, providing evidence for its membrane-modifying activity.

Topics: Aminoisobutyric Acids; Animals; Anti-Bacterial Agents; Ion Channels; Magnetic Resonance Spectroscopy; Mitochondria, Liver; Models, Biological; Oxygen Consumption; Peptaibols; Peptides; Peptides, Cyclic; Protein Conformation; Rats; Thermodynamics; Uncoupling Agents

The crystal structure of the synthetic tetrapeptide, Boc-Hyp-Aib-Aib-Phol, an analogue of the C-terminal tetrapeptide in the antibiotic antiamoebin I, was determined as part of a study of the conformation of peptaibophol antibiotics. The crystals are orthorhombic, space group P212121, with cell parameters a = 16.576 (1) A, b = 17.657 (1) A, c = 10.435 (1) A, V = 3053.9 (2) A3, Z = 4, Dc = 1.163 g.cm-3. The three amino acids from a single turn of a 3 10-helix, stabilized by two intramolecular hydrogen bonds. The Aib residues adopt the usual conformation in the region between the 3 10- and alpha-helices. The terminal hydroxy methyl group of the phenylalaninol residue is disordered. The position of the benzyl side chain of the amino alcohol relative to the backbone corresponds to a conformation also observed in phenylalanine residues.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Models, Molecular; Oligopeptides; Peptaibols; Peptides; Peptides, Cyclic; Protein Conformation; X-Ray Diffraction

Topics: Amino Acid Sequence; Anti-Bacterial Agents; Chemical Phenomena; Chemistry; Hydrolysis; Peptaibols; Peptides, Cyclic

Topics: Anti-Bacterial Agents; Chromatography, Gas; Mass Spectrometry; Peptaibols; Peptides, Cyclic