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LFchimera [id=LFB0154]

Synonym: LF f(265-284)-Lys-f(30-17)

Producer Organism : Native Protein : Production Method :
Cow Lactoferrin (LF) Synthetic
Activity : Antibacterial, antifungal, antiparasitical
Target Organisms :

Gram-positive: Active against Staphylococcus aureus ATCC 25923, Staphylococcus aureus MRSA at 1然, Streptococcus mutans Ingbritt (LC50=1.6 然), Streptococcus sanguis SK4 (LC50=1.5 然).

Gram-negative: Active against Escherichia coli EPEC E2348/69, Escherichia coli MREPEC, Escherichia coli EHEC O157:H7 at 1然, Pseudomonas aeruginosa PAK (LC50=1.2 然), Pseudomonas aeruginosa PAO (LC50=1.5 然), Escherichia coli K12 (LC50=5 然), Helicobacter pylori (MIC=0.2 然), Aggregatibacter actinomycetemcomitans Y4 ATCC 43718 (LC50=5.1 然), Vibrio parahaemolyticus ATCC 17802 (LC50=10 然), Vibrio parahaemolyticus O3:K6 (LC50=10 然), Vibrio cholerae O1 (LC50<10 然), Vibrio cholerae non-O1 (LC50<10 然).

Yeast: Active against Candida albicans 315 ATCC 10231 (no MIC).

Parasite: Active against Entamoeba histolytica (no value), Leishmania donovani (IC50=3.5 然 or LC50=3.5 然), Leishmania pifanoi (IC50=4.5 然 or LC50=6 然).

Description :
Production method: Synthetic.


The innate immunity factor lactoferrin harbours two antimicrobial moieties, lactoferricin (LFB0084) and lactoferrampin (LFB0149), situated in close proximity in the N1 domain of the molecule. Most likely they cooperate in many of the beneficial activities of lactoferrin. To investigate whether chimerization of both peptides forms a functional unit we designed a chimerical structure containing lactoferricin amino acids 17-30 (LFB0030) and lactoferrampin amino acids 265-284 (LFB0131). The bactericidal activity of this LFchimera was found to be drastically stronger than that of the constituent peptides, as was demonstrated by the need for lower dose, shorter incubation time and less ionic strength dependency (Citation: 1).
By electron microscopy, a clear cellular damage was observed in bacteria after treatments with LFchimera and peptides, suggesting that interaction and membrane disruption are probably involved as a mechanism of action (Citation: 2).
Length : 35 Mass (Da): 4 424.13 Common Amino Acids : K
Isolectric Point : 11.85 Net Charge : 11 Absent Amino Acids : HPTVY
Basic Residues : 13 Acidic Residues : 2 Hydrophobic Residues : 11
Polar Residues : 6 Boman Index : -104.81 Hydropathy Index : -1.263
Aliphatic Index : 58.57 Instability Index : 0 Extinction Coefficient : 16500
Absorbance 280nm : 485.29

Wheel representation

Hydrophobicity plot

Red solid plot : values according to the hydrophobicity scale of Kyte and Doolittle (reference paper).
Yellow dashed plot : Experimentally determined hydrophobicity scale for proteins at membrane interfaces(reference paper).
Green dotted-dashed plot : prediction of transmembrane helices (reference paper). In this scale (unlike the others), more negative values reflect greater hydrophobicity.

Citation: 1

Bactericidal activity of LFchimera is stronger and less sensitive to ionic strength than its constituent lactoferricin and lactoferrampin peptides

Cited Entries: LFB0030, LFB0131, LFB0154

Authors:Bolscher, J.G.M., Adao, R., Nazmi, K., van den Keybus, P.A.M., van 't Hof, W., Nieuw Amerongen, A.V., Bastos, M., Veerman, E.C.I.
Journal: Biochimie 2009, 91(1).
CrossRef External Link
Abstract: The innate immunity factor lactoferrin harbours two antimicrobial moieties, lactoferricin and lactoferrampin, situated in close proximity in the N1 domain of the molecule. Most likely they cooperate in many of the beneficial activities of lactoferrin. To investigate whether chimerization of both peptides forms a functional unit we designed a chimerical structure containing lactoferricin amino acids 1730 and lactoferrampin amino acids 265284. The bactericidal activity of this LFchimera was found to be drastically stronger than that of the constituent peptides, as was demonstrated by the need for lower dose, shorter incubation time and less ionic strength dependency. Likewise, strongly enhanced interaction with negatively charged model membranes was found for the LFchimera relative to the constituent peptides. Thus, chimerization of the two antimicrobial peptides resembling their structural orientation in the native molecule strikingly improves their biological activity.
Keywords: antimicrobial peptide; bovine lactoferrin; Lactoferricin; Lactoferrampin; LFchimera; Dsc; Cd
Citation: 2

Bactericidal effect of bovine lactoferrin, LFcin, LFampin and LFchimera on antibiotic-resistant Staphylococcus aureus and Escherichia coli

Cited Entries: LFB0030, LFB0131, LFB0154

Authors:Flores-Villasenor, H., Canizalez-Roman, A., Reyes-Lopez, M., Nazmi, K., de la Garza, M., Zazueta-Beltran, J., Leon-Sicairos, N., Bolscher, J.G.
Journal: BioMetals 2010, 23(3).
Abstract: Increased prevalence of antibiotic-resistant bacteria has become a major threat to the health sector worldwide due to their virulence, limited therapeutic options and distribution in both hospital and community settings. Discovery and development of new agents to combat antibiotic-resistant bacteria is thus needed. This study therefore aimed to evaluate the ability of bovine lactoferrin (LF), peptides from two antimicrobial domains lactoferricin B (LFcin17-30) and lactoferrampin (LFampin265-284) and a chimeric construct (LFchimera) containing both peptides, as potential bactericidal agents against clinical isolates of antibiotic-resistant Staphylococcus aureus and Escherichia coli. Results in kinetics of growth show that LF chimera and peptides inhibited the growth of both bacterial species. By confocal microscopy and flow cytometry it was observed that LF and FITC-labeled peptides are able to interact with these bacteria and cause membrane permeabilization, as monitored by propidium iodide staining, these effects were decreased by preincubation with lipopolysaccharide in E. coli. By electron microscopy, a clear cellular damage was observed in bacteria after treatments with LFchimera and peptides, suggesting that interaction and membrane disruption are probably involved as a mechanism of action. In conclusion, results show that LFchimera, LF and peptides have potential as bactericidal agents in the antibiotic-resistant strains of S. aureus and E. coli and also the work strongly suggest that LFcin17-30 and LFampin265-284 acts synergistically with antibiotics against multidrug resistant EPEC and MRSA in vitro.
Keywords: Chemistry and Materials Science
Citation: 3

Chimerization of lactoferricin and lactoferrampin peptides strongly potentiates the killing activity against Candida albicans

Cited Entries: LFB0030, LFB0131, LFB0154

Authors:Bolscher, J., Nazmi, K., van Marle, J., van t Hof, W., Veerman, E.
Journal: Biochemistry and Cell Biology 2012, 90(3): 11.
Abstract: Bovine lactoferrin harbors 2 antimicrobial sequences (LFcin and LFampin), situated in close proximity in the N1- domain. To mimic their semi parallel configuration we have synthesized a chimeric peptide (LFchimera) in which these sequences are linked in a head-to-head fashion to the a- and 3-amino group, respectively, of a single lysine. In line with previously described bactericidal effects, this peptide was also a stronger candidacidal agent than the antimicrobial peptides LFcin1730 and LFampin265284, or a combination of these 2. Conditions that strongly reduced the candidacidal activities of LFcin1730 and LFampin265284, such as high ionic strength and energy depletion, had little influence on the activity of LFchimera. Freeze-fracture electron microscopy showed that LFchimera severely affected the membrane morphology, resulting in disintegration of the membrane bilayer and in an efflux of small and high molecular weight molecules such as ATP and proteins. The differential effects displayed by the chimeric peptide and a mixture of its constituent peptides clearly demonstrate the synergistic effect of linking these peptides in a fashion that allows a similar spatial arrangement as in the parent protein, suggesting that in bovine lactoferrrin the corresponding fragments act in concert in its candidacidal activity
Keywords: Candida albicans, yeast, pseudohyphae, lactoferricin, lactoferrampin, LFchimera
Citation: 4

Enhanced leishmanicidal activity of cryptopeptide chimeras from the active N1 domain of bovine lactoferrin

Cited Entries: LFB0030, LFB0131, LFB0154, LFB0173, LFB0174, LFB0175, LFB0176, LFB0177

Authors:Silva, T., Abengozar, M. A., Fernandez-Reyes, M., Andreu, D., Nazmi, K., Bolscher, J. G. M., Bastos, M., Rivas, L.
Journal: Amino acids 2012,.
Abstract: Two antimicrobial cryptopeptides from the N1 domain of bovine lactoferrin, lactoferricin (LFcin1730) and lactoferrampin (LFampin265284), together with a hybrid version (LFchimera), were tested against the protozoan parasite Leishmania. All peptides were leishmanicidal against Leishmania donovani promastigotes, and LFchimera showed a significantly higher activity over its two composing moieties. Besides, it was the only peptide active on Leishmania pifanoi axenic amastigotes, already showing activity below 10 lM. To investigate their leishmanicidal mechanism, promastigote membrane permeabilization was assessed by decrease of free ATP levels in living parasites, entrance of the vital dye SYTOX Green (MW = 600 Da) and confocal and transmission electron microscopy. The peptides induced plasma membrane permeabilization and bioenergetic collapse of the parasites. To further clarify the structural traits underlying the increased leishmanicidal activity of LFchimera, the activity of several analogues was assessed. Results revealed that the high activity of these hybrid peptides seems to be related to the order and sequence orientation of the two cryptopeptide moieties, rather than to their particular linkage through an additional lysine, as in the initial LFchimera. The incorporation of both antimicrobial cryptopeptide motifs into a single linear sequence facilitates chemical synthesis and should help in the potential clinical application of these optimized analogues.
Keywords: Leishmania; Leishmanicidal activity; Antimicrobial peptides; Membrane permeabilization; Lactoferricin; Lactoferrampin; LFchimera
Citation: 5

Bactericidal effect of lactoferrin and lactoferrin chimera against halophilic Vibrio parahaemolyticus

Cited Entries: LFB0030, LFB0131, LFB0154

Authors:Leon-Sicairosa, N., Canizalez-Romanb, A., de la Garzac, M., Reyes-Lopez, M., Zazueta-Beltrand, J., Nazmif, K., Gomez-Gile, B., Bolscher, J. G.
Journal: Biochimie 2009, 91(1): 8.
Abstract: Infections caused by Vibrio parahaemolyticus, an halophilic member of the genus Vibrio, have increased globally in the last 5 years. Diarrhea caused by V. parahaemolyticus results from eating raw or undercooked seafood. The aim of this work was to investigate whether lactoferrin and some lactoferrin-peptides have bactericidal activity against Vibrio parahaemolyticus ATCC 17802, the pandemic strain O3:K6, and the multidrug resistant isolate 727, as well as against Vibrio cholerae strains O1 and non-O1. Whereas both peptides lactoferricin (1730) and lactoferrampin (265284) did not have bactericidal activity, 40 μM of lactoferrin chimera (a fusion of the two peptides) inhibited the growth of all Vibrio tested to the same extent as the antibiotic gentamicin. The cidal effect of LFchimera showed a clear concentration response in contrast to bovine lactoferrin which showed higher inhibition at 10 μM than at 40 μM. FITC-labeled LFchimera bound to the bacterial membranes. Moreover LFchimera permeabilized bacterial cells and membranes were seriously damaged. Finally, in experiments with the multidrug resistant isolate 727, sub-lethal doses of LFchimera strongly reduced the concentrations of ampicillin, gentamicin or kanamicin needed to reach more than 95% growth inhibition, suggesting synergistic effects. These data indicate that LFchimera is a potential candidate to combat the multidrug resistant pathogenic Vibrio species.
Keywords: Lactoferrin; Lactoferricin; Lactoferrampin; LFchimera; Vibrio parahaemolyticus
Citation: 6

Microbicidal effect of the lactoferrin peptides Lactoferricin1730, Lactoferrampin265284, and Lactoferrin chimera on the parasite Entamoeba histolytica

Cited Entries: LFB0030, LFB0131, LFB0154

Authors:Lopez-Soto, F., Leon-Sicairos, N., Nazmi, K., Bolscher, J. G., de la Garza, M.
Journal: Biometals 2010, 23: 6.
Abstract: Entamoeba histolytica is a parasitic protozoan that produces amoebiasis, an intestinal disease characterized by ulcerative colitis and dysentery. In some cases, trophozoites can travel to the liver leading to hepatic abscesses and death. Recently, lactoferrin and lactoferricin B have been shown to be amoebicidal in axenic cultures. The aim of this work was to determine whether the lactoferrin-peptides lactoferricin amino acids 1730, lactoferrampin amino acids 265284, and lactoferrin chimera which is a fusion product of the two peptides, are capable of producing a microbicidal effect to trophozoites of E. histolytica. We evaluated the killing effect of these peptides in growth kinetics carried out in axenic culture medium to which different concentrations of peptides were added. At 50 lM of peptide concentration, lactoferricin and lactoferrampin had a moderate amoebicidal effect, since a 4550% of trophozoites remained viable at 24 h culture. However, at 50 lM of the lactoferrin chimera 75% amoeba were killed whereas at 100 lM all cells died. These data indicate that of lactoferrin-peptides mainly the chimera have amoebicidal activity in a time- and concentration-dependent manner. The lactoferrin-peptides might be useful as therapeutic agents against amoebiasis and thereby diminish the use of metronidazole, which is extremely toxic for the host.
Keywords: Entamoeba histolytica; Amoebicidal effect, LFcin; LFampin; LFchimera

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