Академический Документы
Профессиональный Документы
Культура Документы
Family of Inducible antimicrobial oligopeptides ; small proteins of about 35 amino acid residues active against both Gram-positive and Gram-negative bacteria
These peptides play an important role in the offense and defense systems of different organisms .
Are an evolutionarily conserved component of the innate immune response.
COMMON CHARACTERISTICS
- Amino acid composition - Amphipathicity - Cationic charge - Size - their synthesis is of type ribosomal All these characteristics allow them to attach to and insert into membrane bilayers to form pores .
CECROPIN
Inducible antimicrobial oligopeptide discovered for the first time in the hemolymph of Lepidoptera
CECROPINS
Linear peptides that are structured to -helix in hydrophobic ambient. They assume random structure in solution.
CECROPINS
They perform host defense functions and often act in synergy, providing rapid non-specific defence against invading micro-organisms . Most of these natural broad-spectrum peptide antibiotics have limited cytotoxicity to human cells and are being developed as therapeutics against pathogens resistant to classical antibiotics.
type A, B and D
Cecropins A, B and D :
- Are linear peptides - Are basic peptides - Consist of 35-37 residues that show 62-65% homology in the primary sequence
CECROPIN- A : DATA
Formula : C184H313N53O46
KWKLFKKIEKVGQNIRDGIIKAGPAVAVVGQATQIAK-NH2
CECROPIN- A
Isolated from the silkworm Hyalophora cecropia . Composed of 37 amino acids. Kills bacteria by dissipating transmembrane electrochemical ion-gradients. Cecropin A induces changes in gene expression profiles in Escherichia coli exposed to sub lethal doses, apart from any lethal effects caused by membrane perturbation.
CECROPIN- B : DATA
Linear and cationic
KWKVFKKIEKMGRNIRNGIVKAGPAIAVLGEAKAL-NH2
CECROPIN- B
Isolated from the giant silkmoth, Hyalophora cecropia. It enhances the activities of betalactams in experimental rat models of Gram-negative septic shock. Cecropin B has been shown to reduce significantly the lethality of Escherichia coli and plasma endotoxin levels in a rat model of septic shock . Can affect host cell to promote wound repair.
Cecropin D :
- It appears in the hemolymph at much later times after bacterial infection than Cecropin A or Cecropin B . - It possesses potent antibacterial activities against a panel of Gram positive and negative bacteria without haemolytic activity against human red blood cells. - These peptides are more active against Bacillus megaterium than Cecropin A.
Some time later, many other Cecropine were isolated from organisms other than Hyalophora cecropia . Cecropins isolated from other insects have been given various names bactericidin, lepidopteran, sarcotoxin, etc. All of these peptides are structurally related.
Cecropin- AD :
The peptide is a hybrid analog consisting of cecropin A-(1-11) and D-(12-37). The hybrid protein is 5-55 times as active as cecropin D against some bacteria and slightly more active than cecropin A. It is 6 times more active than cecropin A against Bacillus subtilis Bs11. It has reported that Cecropin AD promotes cell death by apoptosis in Nasopharyngeal carcinoma cells.
Cecropin P1 :
- It has been isolated originally as anti Escherichia coli factor from pig intestines . - This peptide is not of mammalian origin, but is
produced by the pig intestinal parasitic nematode, Ascaris suum, and human Ascaris lumbricoides - Is bactericidal against a wide range of microorganisms: Aeromonas hydrophila, Bacillus subtilis , Escherichia coli; Klebsiella pneumoniae, Micrococcus luteus, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pneumocystis carinii, Saccharomyces cerevisiae .
CECROPIN- XJ:
Is extremely heat-stable and tolerates extreme acidic, basic, and high salt environments. Has the ability to kill ampicillin- resistant Staphylococcus aureus and Staphylococcus enterica . It is also resistant to 24 hours digestion by artificial gastric juice.
Function of Cecropin
The site of action of the antimicrobial activity is the cell membrane. Cecropin- A folds into final structure while superficially adsorbed to a membrane surface. Interactions with deeper hydrophobic regions of the bilayer appear to be unnecessary for folding. Its conformation is predominantly -helical with some structure.
To investigate the nature of its membrane activity, the interactions of cecropin with synthetic lipid vesicles, bacteria and planar lipid membranes were investigated using various techniques .
Action of Cecropins
Just in vesicles, the action of cecropin A is concentration dependent, forming : - ion channels at low peptide to lipid ratios - pores ,large enough to pass probe molecules, at higher peptide to lipid ratios.
The bactericidal action of Cecropins is correlated with the permeabilization of the membrane, that is , infact, always followed by irreversible cytolysis and death of the bacteria.
Action of Cecropins
Many of these peptides directly act by altering the membrane of target cells: the interaction of these molecules with some chiral receptor membrane seems excluded, since analogues of these peptides consisting of all enantiomers D are active as the natural ones.
Toroidal Model
The peptide binds to the membrane and when it reaches the critical value pep/lip 1:20 changes its orientation
The toroidal pore is formed when the alpha helices of Cecropins intercalate in the lipid bilayer
Barrel Stave
Toroidal
All the Cecropins cause the formation of a toroidal pore in the phospholipid bilayer
Action of Cecropin- P1
Interaction with :
Gram +
The peptide is probably attracted by the acids teicoici and teicuronici and other anionic groups which are located externally to the layer of peptidoglycan.
Gram
Initially the peptide interacts with the polyanionic molecules of lipopolysaccharide of the outer membrane and is then able to make it permeable or to be received inside.
Methods
The antiproliferative and cytotoxic potential of the Cecropins A and B was quantified by assays in four bladder cancer cell lines as well as in murine and human fibroblast cell lines. Scanning electron microscopy (SEM) was performed to visualize the morphological changes induced by Cecropin A and B in bladder tumor cells and benign fibroblasts.
Results
1. Cecropin A and B inhibit bladder cancer cell proliferation and viability in a dosedependent fashion. 2. Benign fibroblasts were significantly less or not at all susceptible to Cecropin A and B.
3. Both Cecropins induced an increase in LDH release from bladder tumor cells whereas benign fibroblasts were not affected.
4. Transfection of human bladder cancer cells with Cecropin genes reduces their tumorigenicity in nude mouse models too.
Conclusions
Cecropin A and B exert selective cytotoxic and antiproliferative efficacy in bladder cancer cells while sparing targets of benign murine or human fibroblast origin. Both peptides may offer novel therapeutic strategies for the treatment of bladder cancer with limited cytotoxic effects on benign cells.
Leishmaniasis
Disease caused by protozoan parasites that belong to the genus Leishmania and is transmitted by the bite of certain species of sand fly.
The hibrid Cecropin- Melittin most active against Leishmaniasis is CA(1-8)M(1-18) A 4 M the proliferation of promastigotes is inhibited up to 95% .
The action of the peptide in Leishmaniasis is very rapid. At a concentration of 2.5 pM, the 90% of the final effect is reached in the first 5 minutes of incubation.
Factors of interaction
Temperature : the maximum inhibition of
lipophosphoglican ( LPG ) is one of major components of promastigotes membrane, and it binds Ca2 + cations. If we treat the cells with EDTA (Ethylenediaminetetraacetic acid ) , which sequesters these cations, you can observe a strong awareness of promastigotes to the peptide .
Alterations caused
Damage of the flagellar pocket , that after the treatment is devoid of microtubules . Increase of the permeability of mebrane: it causes the accumulation of proline that affects both the pH gradient and the membrane potential , the two components involved in the maintenance of energetic capacity by translocation of protons.
Cecropins are especially promising candidates for anticancer therapy in humans because they demonstrate several unique features:
1. Their selectivity for malignant cells and their potentially pronounced lytic activity against high-grade tumor cells allow for an optimal therapy in vivo with low therapeutic concentrations and limited side effects .
2. The anti-tumoral effect of Cecropins has been hypothesized to be unaffected by the multidrug resistance (MDR) phenotype observed in many cancer types. 3. Classic chemotherapeutic agents such as mitomycin which are widely used for intravesical instillation are highly unstable in urine. In contrast, Cecropins are largely resistent against serum and urine proteolysis because of their specific biochemical structures, rendering them ideal candidates for intravesical tumor therapy .