Peptide having cell membrane penetrating activity

Abstract

Provided are transmembrane complexes that contain a protein transduction domain (PTD) from the N-terminus of IgE-dependent histamine-releasing factor (HRF) and a target substance that is to be delivered into a cell. Also provided are nucleic acid molecules encoding the transmembrane complex, and methods of delivering the target substance into a cell interior by contacting the transmembrane complex with a cell. Also provided are transfection kits containing the PTD and the target substance.

1 . A transmembrane complex, comprising:
 a) a protein transduction domain (PTD) that comprises all or a portion of the N-terminus of IgE-dependent histamine-releasing factor (HRF), wherein the PTD:
 has cell membrane penetrating activity; 
 the portion contains at least 5 amino acids; 
 the PTD contains 5-15 amino acids; and 
 the N-terminus of HRF consists of the sequence MIIYRDLISH; and. 
   b) a target substance, whereby the transmembrane complex is not HRF.   
     
     
         2 . A transmembrane complex of  claim 1 , consisting of the PTD and the target substance. 
     
     
         3 . A transmembrane complex of  claim 1 , wherein the PTD contains 8-12 amino acids. 
     
     
         4 . The transmembrane complex of  claim 1 , wherein the PTD comprises a sequence of amino acids selected from among SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 12; SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 22, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53 and SEQ ID NO: 54. 
     
     
         5 . The transmembrane complex of  claim 1 , wherein the transmembrane complex is a fusion protein and the fusion protein is prepared by connecting the target substance to linker, and reacting with the peptide having cell membrane penetrating activity to form linkage. 
     
     
         6 . The transmembrane complex of  claim 1 , wherein the target substance is selected from among a nucleic acid, a drug, a chemical compound, a carbohydrate, a lipid, a glycolipid, an enzyme, a regulating factor, a growth factor and an antibody. 
     
     
         7 . A transfection kit, comprising:
 a) a first composition containing a protein transduction domain (PTD) that comprises all or a portion of the N-terminus of IgE-dependent histamine-releasing factor (HRF), wherein the PTD:
 has cell membrane penetrating activity; 
 the portion contains at least 5 amino acids; 
 the PTD contains 5-15 amino acids; and 
 the N-terminus of HRF consists of the sequence MIIYRDLISH; and 
   b) a second composition comprising a target substance.   
     
     
         8 . The transfection kit of  claim 7 , further comprising a binding factor for linking the PTD to the target substance. 
     
     
         9 . A method for delivering a target substance into a cell interior, comprising administering a complex of  claim 1  to a subject, whereby the complex is delivered into the cell. 
     
     
         10 . The method of  claim 9 , wherein the PTD comprises a sequence of amino acids selected from among SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 12; SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 22, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53 and SEQ ID NO: 54. 
     
     
         11 . The method of  claim 9 , wherein the target substance is selected from among a nucleic acid, a drug, a chemical compound, a carbohydrate, a lipid, a glycolipid, an enzyme, a regulating factor, a growth factor and an antibody. 
     
     
         12 . A nucleic acid molecule encoding a transmembrane complex, comprising a sequence of nucleotides that encodes a protein transduction domain (PTD) linked to target substance, wherein:
 a) the PTD:
 has cell membrane penetrating activity; 
 the portion contains at least 5 amino acids; 
 the PTD contains 5-15 amino acids; and 
 the N-terminus of HRF consists of the sequence MIIYRDLISH; and. 
   b) the target substance is a nucleic acid molecule encoding a polypeptide for delivery, provided that the encoded complex is not IgE-dependent histamine-releasing factor (HRF).   
     
     
         13 . The nucleic acid molecule of  claim 12 , wherein the PTD comprises a sequence of amino acids selected from among SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 12; SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 22, SEQ 1D NO: 26, SEQ ID NO: 27, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53 and SEQ ID NO: 54. 
     
     
         14 . The nucleic acid molecule of  claim 12 , wherein the target substance is an enzyme, a regulating factor, a growth factor or an antibody. 
     
     
         15 . The nucleic acid molecule of  claim 12 , wherein the PTD is encoded by a sequence of nucleotides selected from among one of SEQ ID NOS: 17, 18 and 55-81. 
     
     
         16 . A method of delivering a target substance into a cell interior, comprising
 a) linking a protein transduction domain (PTD) to a target substance by physically/chemically covalent bond or non-covalent bond or by mediator in incorporated or fused form to form a transmembrane complex; and   b) administering the transmembrane complex to a subject,   wherein the protein transduction domain (PTD) comprises all or a portion of the N-terminus of IgE-dependent histamine-releasing factor (HRF),   wherein the PTD:
 has cell membrane penetrating activity; 
 the portion contains at least 5 amino acids; 
 the PTD contains 5-15 amino acids; and 
 the N-terminus of HRF consists of the sequence MIIYRDLISH. 
   
     
     
         17 . The method of  claim 16 , wherein the PTD comprises a sequence of amino acids selected from among SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO:4, SEQ ID NO: 12; SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO: 22, SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 31, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 34, SEQ ID NO: 35, SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 43, SEQ ID NO: 44, SEQ ID NO: 45, SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53 and SEQ ID NO: 54. 
     
     
         18 . The method of  claim 16 , wherein the target substance is selected from among a nucleic acid, a drug, a chemical compound, a carbohydrate, a lipid, a glycolipid, an enzyme, a regulating factor, a growth factor and an antibody. 
     
     
         19 . The transmembrane complex of  claim 1  that is a fusion protein, comprising the PTD linked to the target substance directly or via a linker. 
     
     
         20 . The transmembrane complex of  claim 19 , wherein the PTD is linked to the target substance via a linker.