Materials for the production of nanometer structures and use thereof

Abstract

The present invention pertains to nanostructures, i.e., nanometer sized structures useful in the construction of microscopic and macroscopic structures. In particular, the present invention pertains to nanostructures based on bacteriophage T4 tail fiber proteins and variants thereof.

What is claimed is:  
     
         1 . An isolated polypeptide consisting essentially of the gp37 tail fiber protein of bacteriophage T4 lacking amino acids 99-496 (SEQ ID NO: 6) when numbered from the amino terminus, wherein said polypeptide has the capability to form dimers and interact with the P36 protein oligomer of bacteriophage T4.  
     
     
         2 . An isolated polypeptide consisting essentially of a fusion protein between the gp36 and gp37 proteins of bacteriophage T4, wherein amino acid residues 1-242 of gp37 (SEQ ID NO: 6) are fused in proper reading frame to amino acid residues 118-221 of gp36 (SEQ ID NO: 5).  
     
     
         3 . The polypeptide of  claim 2  wherein a plurality of carboxy termini of said polypeptide have the capability of interacting with the amino terminus of the P37 protein oligomer of bacteriophage T4 and to form an attached oligomer and the amino termini of the oligomer of said polypeptide have the capability of interacting with the carboxy termini of gp36 polypeptides of bacteriophage T4.  
     
     
         4 . An isolated polypeptide oligomer consisting essentially of two gp37 polypeptides of bacteriophage T4, wherein the amino termini of said oligomer lack the capability of interacting with the carboxy termini of gp36 polypeptides of bacteriophage T4.  
     
     
         5 . An isolated polypeptide oligomer consisting essentially of the P37 protein of bacteriophage T4, wherein the amino termini of said oligomer lack the capability of interacting with the carboxy termini of gp36 polypeptides of bacteriophage T4.  
     
     
         6 . An isolated polypeptide consisting essentially of a variant of the gp36 protein of bacteriophage T4, wherein said polypeptide lacks the capability of interacting with the amino terminus of the P37 protein oligomer of bacteriophage T4.  
     
     
         7 . An isolated polypeptide consisting essentially of a fusion protein between the gp36 and gp34 proteins of bacteriophage T4, wherein amino acid residues 1-73 of gp36 (SEQ ID NO: 5) are fused in proper reading frame amino-terminal to amino acid residues 866-1289 of gp34 (SEQ ID NO: 2).  
     
     
         8 . An oligomer of the polypeptide of  claim 7 , wherein the amino termini of said dimer have the capability of interacting with the gp35 protein of bacteriophage T4.  
     
     
         9 . An isolated polypeptide consisting essentially of a variant of the gp35 protein of bacteriophage T4, wherein said polypeptide forms an angle of less than about 125° when combined with the P34 and P36-P37 protein oligomers of bacteriophage T4, under conditions wherein the wild-type gp35 protein forms an angle of 137° when combined with said oligomers.  
     
     
         10 . An isolated polypeptide consisting essentially of a variant of the gp35 protein of bacteriophage T4, wherein said polypeptide forms an angle of more than about 145° when combined with the P34 and P36-P37 protein oligomers of bacteriophage T4, under conditions wherein the wild-type gp35 protein forms an angle of 137° when combined with said oligomers.  
     
     
         11 . An isolated polypeptide consisting essentially of a variant of the gp35 protein of bacteriophage T4, wherein the interaction of said polypeptide with the P34 protein oligomer of bacteriophage T4 is unstable at temperatures between about 40° C. and about 60° C.  
     
     
         12 . An isolated polypeptide oligomer consisting essentially of a variant of the P37 protein of bacteriophage T4, wherein the interaction of said oligomer with the P36 protein oligomer of bacteriophage T4 is unstable at temperatures between about 40° C. and about 60° C.  
     
     
         13 . An isolated polypeptide oligomer consisting essentially of a variant of the P37 protein of bacteriophage T4, wherein the carboxy-terminal domain of said oligomer is modified so as to confer the ability of the entire polypeptide to bind specifically to an immobilized ligand.  
     
     
         14 . The polypeptide of  claim 13 , wherein said ligand is selected from the group consisting of biotin, immunoglobulin, or divalent metal ions.  
     
     
         15 . A nanostructure comprising a plurality of fusion proteins, said fusion proteins comprising a first portion consisting of at least the first 10 N-terminal amino acids of a tail fiber protein fused via a peptide bond to a second portion consisting of at least the last 10 C-terminal amino acids of a second tail fiber protein, wherein the tail fiber proteins are selected from the group consisting of gp34, gp35, gp36, and gp37 proteins of a T-even-like bacteriophage, wherein the first and second tail fiber proteins are the same or different.  
     
     
         16 . The nanostructure of  claim 15 , wherein the first and second tail fiber proteins are different.  
     
     
         17 . The nanostructure of  claim 15 , which further comprises a molecule that can self-assemble into a dimer or trimer, fused to at least a 10 amino acid portion of a T-even-like tail fiber protein.  
     
     
         18 . The nanostructure of  claim 17 , wherein the molecule has the structure of a leucine zipper.  
     
     
         19 . The nanostructure of  claim 15 , wherein said nanostructure comprises a linear one-dimensional rod.  
     
     
         20 . The nanostructure of  claim 15 , wherein said nanostructure comprises a polygon.  
     
     
         21 . The nanostructure of  claim 15 , wherein said nanostructure comprises a three-dimensional cage or solid.  
     
     
         22 . The nanostructure of  claim 15 , wherein said nanostructure comprises a two-dimensional open or closed sheet.  
     
     
         23 . An isolated fusion protein consisting essentially of a portion of a gp37 protein of a T-even-like bacteriophage consisting of at least the first 10-60 N-terminal amino acids of the gp37 protein fused to a second portion of a gp36 protein of a T-even-like bacteriophage consisting of at least the last 10-60 C-terminal amino acids of the gp36 protein.  
     
     
         24 . An isolated fusion protein consisting essentially of a portion of a gp37 protein of a T-even-like bacteriophage consisting of at least the first 10 N-terminal amino acids of the gp37 protein fused to a second portion of a gp36 protein of a T-even-like bacteriophage consisting of at least the last 10 C-terminal amino acids of the gp36 protein.  
     
     
         25 . An isolated fusion protein consisting essentially of a portion of a gp37 protein of a T-even-like bacteriophage consisting of at least the first 20 N-terminal amino acids of the gp37 protein fused to a second portion of a gp36 protein of a T-even-like bacteriophage consisting of at least the last 20 C-terminal amino acids of the gp36 protein.  
     
     
         26 . An isolated fusion protein consisting essentially of a portion of a gp36 protein of a T-even-like bacteriophage consisting of at least the first 10-60 N-terminal amino acids of the gp36 protein fused to a second portion of a gp34 protein of a T-even-like bacteriophage consisting of at least the last 10-60 C-terminal amino acids of the gp34 protein.  
     
     
         27 . An isolated protein comprising at least 20 contiguous amino acids of the gp37, gp36, or gp34 protein of a T-even-like bacteriophage, and lacking at least 5 amino acids of the amino- or carboxy-terminus of the protein.  
     
     
         28 . An isolated DNA encoding the polypeptide of  claim 1 .  
     
     
         29 . An isolated DNA encoding the polypeptide of  claim 2 .  
     
     
         30 . An isolated DNA encoding the polypeptide of  claim 4 .  
     
     
         31 . An isolated DNA encoding the polypeptide of  claim 5 .  
     
     
         32 . An isolated DNA encoding the polypeptide of  claim 6 .  
     
     
         33 . An isolated DNA encoding the polypeptide of  claim 7 .  
     
     
         34 . An isolated DNA encoding the polypeptide of  claim 9 .  
     
     
         35 . An isolated DNA encoding the polypeptide of  claim 10 .  
     
     
         36 . An isolated DNA encoding the polypeptide of  claim 11 .  
     
     
         37 . An isolated DNA encoding the polypeptide of  claim 12 .  
     
     
         38 . An isolated DNA encoding the polypeptide of  claim 13 .  
     
     
         39 . An isolated DNA encoding the protein of  claim 23 .  
     
     
         40 . An isolated DNA encoding the protein of  claim 25 .  
     
     
         41 . An isolated DNA encoding the protein of  claim 26 .  
     
     
         42 . An isolated DNA encoding the protein of  claim 27 .  
     
     
         43 . A method for making a polygonal nanostructure comprising contacting the protein of  claim 26  with purified gp35 proteins of a T-even-like bacteriophage.  
     
     
         44 . A method for making a nanostructure comprising contacting a plurality of the proteins of  claim 23  with each other.  
     
     
         45 . A kit comprising in one or more containers the fusion protein of  claim 23 .  
     
     
         46 . A kit comprising in one or more containers the fusion protein of  claim 25 .  
     
     
         47 . A kit comprising in one or more containers the fusion protein of  claim 26 .  
     
     
         48 . A kit comprising in one or more containers the fusion protein of  claim 26 , and an isolated gp35 protein of a T-even-like bacteriophage.  
     
     
         49 . The protein of  claim 23  wherein the T-even-like bacteriophage is T4.  
     
     
         50 . The protein of  claim 26  wherein the T-even-like bacteriophage is T4.  
     
     
         51 . An isolated polypeptide consisting essentially of a variant of the gp36 protein of bacteriophage T4, wherein the interaction of said polypeptide with the P37 protein oligomer of bacteriophage T4 is unstable at temperatures between about 40° C. and about 60° C.  
     
     
         52 . An isolated polypeptide consisting essentially of a variant of the gp36 protein of bacteriophage T4, wherein the interaction of said polypeptide with the gp35 protein of bacteriophage T4 is unstable at temperatures between about 40° C. and about 60° C.  
     
     
         53 . An isolated polypeptide consisting essentially of a variant of the gp34 protein of bacteriophage T4, wherein the interaction of said polypeptide with the gp35 protein of bacteriophage T4 is unstable at temperatures between about 40° C. and about 60° C.