Methods of using helical peptides in plants

Abstract

Novel synthetic lytic and proliferative peptides were designed and constructed to encompass the structural features associated with lytic and proliferative activity. These compounds, along with the human β fibrin signal peptide share structural and functional properties of the known lytic peptides. These peptides are effective agents in the treatment of microbial infections including gram negative and gram positive bacteria, fungus, virus, yeast, and protozoa, in the lysis of cancer cells, and in the proliferation of fibroblasts and lymphocytes. Additional functions include synergy and use as general adjuvants and in the enhancement of wound healing.

1 . A peptide comprising: 
 (a) an α-helical conformation of approximately 15 to 40 amino acids;    (b) one or more sets of 18 sequential amino acids, each amino acid spaced 20 apart about the axis of the helix, exhibiting approximately 60-100% aligned amphipathy;    (c) four or more positively charged amino acids;    (d) where the amino acid at position 19 is not glutamate or aspartate, and the amino acid at position 23 or 24 is not proline; and    (e) where no four consecutive amino acids are of the sequence: lysine-arginine-lysine-arginine.    
     
     
         2 . The peptide of  claim 1 , wherein the peptide is comprised of less than 20 amino acids.  
     
     
         3 . The peptide of  claim 1 , wherein only one set of 18 sequential amino acids exhibit aligned amphipathy.  
     
     
         4 . The peptide of  claim 3 , wherein the set of 18 sequential amino acids exhibiting amphipathy is located at the carboxy terminal half of the peptide.  
     
     
         5 . The peptide of  claim 3 , wherein the set of 18 sequential amino acids exhibiting amphipathy is located at the amino terminal half of the peptide.  
     
     
         6 . The peptide of  claim 1 , wherein said peptide is comprised of combinations of 3 to 5 amino acids.  
     
     
         7 . The peptide of  claim 6 , wherein said amino acids are selected from the group comprising: alinine, lucine, phenyalanine, lysine, and glutamine.  
     
     
         8 . The polypeptide of  claim 2 , wherein said polypeptide has the amino acid sequence: 
 LPKWKVFKKIEKVGRNIRNGIVKAGPAIAVLGEAKALG    
     
     
         9 . The polypeptide of  claim 2 , wherein: 
 said polypeptide has the amino acid sequence:    MKRMVSWSFKKLKTMKKLLLLLLCVFLVKS    
     
     
         10 . The polypeptide of  claim 2 , wherein: 
 said polypeptide has the amino acid sequence:    MKRMVSWSFRKLKTMKRLLLLLLCVFLVKS    
     
     
         11 . The polypeptide of  claim 2 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKKLAKKLKKLAKKLAKLALAL    
     
     
         12 . The polypeptide of  claim 2 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKKLAKKLKKLAKKLAKLALALKALALKAL    
     
     
         13 . The polypeptide of  claim 2 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKKLAKKLKKLAKKLAKLALALKALALKALALKALAL    
     
     
         14 . The polypeptide of  claim 3 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FALALKALKKALKKLKKALKKAL    
     
     
         15 . The polypeptide of  claim 3 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKLALAKLALALKALKKALKKLKKALKKAL    
     
     
         16 . The polypeptide of  claim 3 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FALAKLALAKLALAKLALALKALKKALKKLKKALKKAL    
     
     
         17 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKKLAKLAKKLAKLAKKLAKKL    
     
     
         18 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKKLAKKLKKLAKKLAKLAKKL    
     
     
         19 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKKLAKKLKKLAKKLAKLAKKLAKKLKKLA    
     
     
         20 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    FAKKLAKKLKKLAKKLAKLAKKLAKKLKKLAKKLAKLA    
     
     
         21 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    QFAQKLAKLAQQLAKKLQQLAKK    
     
     
         22 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    QLQAKLKAQLQAKLKAQLQAKLK    
     
     
         23 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    EFAEKLAKLAEELAKKLEELAKK    
     
     
         24 . The polypeptide of  claim 1 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    LKKLAKLAKKLAKLAKKLAKK    
     
     
         25 . The polypeptide of  claim 4 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    MPKWKVFKKIEKVGRNIRN    
     
     
         26 . The polypeptide of  claim 4 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    MPRWRLFRRIDRVGKQIKQ    
     
     
         27 . The polypeptide of  claim 4 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    MPKEKVFLKIEKMGRNIRN    
     
     
         28 . The polypeptide of  claim 4 , wherein: 
 said polypeptide is comprised of the amino acid sequence:    MKRMVSWSFKKLKTMKKLL    
     
     
         29 . A method of killing microbial cells comprising: 
 contacting said pathogenic microorganisms with an effective dose of a therapeutic composition comprising the polypeptide of  claim 1 .    
     
     
         30 . The method of  claim 29  wherein: 
 said contacting is with a therapeutic composition comprising human beta fibrin signal peptide.  
 
     
     
         31 . The method of  claim 30  wherein: 
 said pathogenic microorganisms are selected from the group comprising: bacteria, fungi, yeast and protozoa.  
 
     
     
         32 . The method of  claim 31  wherein: 
 said contacting is with a therapeutic composition comprising human beta fibrin signal peptide.  
 
     
     
         33 . The method of  claim 32  wherein: 
 said bacteria are selected from gram positive and gram negative bacteria.  
 
     
     
         34 . The method of  claim 33  wherein: 
 said contacting is with a therapeutic composition comprising human beta fibrin signal peptide.  
 
     
     
         35 . A method of treating microbial infection in plants and animals comprising: 
 administering to the plants or animals an effective dose of a therapeutic composition comprising the polypeptide of  claim 1 .    
     
     
         36 . The method of  claim 35 , wherein: 
 said administering is with a therapeutic composition comprising human beta fibrin signal peptide.    
     
     
         37 . The method of  claim 35  wherein: 
 said microbial infection is selected from the group comprising: bacterial, viral, fungal, yeast or protozoan infection.  
 
     
     
         38 . The method of  claim 35  wherein: 
 said administering is by means selected from the group comprising: topical application, injection and infusion.  
 
     
     
         39 . A method for the treatment of cancer in animals comprising: 
 administering to said animals an effective dose of a therapeutically active composition comprising the polypeptide of  claim 1 .    
     
     
         40 . The method of  claim 39  wherein: 
 said administering is with a theraupeutic composition comprising human beta fibrin signal peptide.  
 
     
     
         41 . A method for the prevention of a pathogenic disease in plants and animals comprising: 
 incorporating into the plant or animal's genome one or more genes encoding the polypeptide of  claim 1 .    
     
     
         42 . The method of  claim 41 , wherein: 
 said administering is with a therapeutic composition comprising human beta fibrin signal peptide.    
     
     
         43 . A method for the proliferation of cell growth comprising: 
 contacting the cells with an effective dose of a therapeutic composition comprising the polypeptide of  claim 1 .    
     
     
         44 . The method of  claim 43  wherein: 
 said contacting is with a therapeutic composition comprising human beta fibrin signal peptide.  
 
     
     
         45 . The method of  claim 43 , wherein: 
 said cells are selected from the group comprising white blood cells and fibroblasts.    
     
     
         46 . A method for the healing of wounds comprising: 
 administering to the wound an effective dose of a therapeutic composition comprising the polypeptide of  claim 1 .    
     
     
         47 . The method of  claim 46 , wherein: 
 said administering is with a therapeutic composition comprising human beta fibrin signal peptide.    
     
     
         48 . A method to facilitate the action of therapeutic agents comprising: 
 administering said therapeutic agents in a composition comprising an effective dose of the polypeptide of  claim 1 .    
     
     
         49 . The method of  claim 48 , wherein: 
 said administering is with a therapeutic composition comprising human beta fibrin signal peptide.