Metal binding proteins and associated methods

Abstract

Metal binding proteins, associated compositions and methods for their production and use are disclosed. The metal binding proteins included have amino acid sequences analogous to at least one metal binding protein, and conservative amino acid substitutions thereof from a brine shrimp ( Artemia ). Also provided are the associated nucleic acid sequences encoding metal binding proteins.

1 . A substantially purified metal binding protein having an amino acid sequence analogous to at least one metal binding protein from a brine shrimp ( Artemia ). 
     
     
         2 . The substantially purified metal binding protein of  claim 1  including an amino acid sequence selected from the group consisting of: 
       
         
           
                 
               
                   [SEQ ID NO: 2] 
                 
                 
                 
               
                   (1) MET ASP CYS CYS LYS ASN GLY CYS THR CYS ALA 
                     
                 
                     
                 
                   PRO ASN CYS LYS CYS ALA LYS ASP CYS LYS CYS CYS 
                 
                     
                 
                   LYS GLY CYS GLU CYS LYS SER ASN PRO GLU CYS LYS 
                 
                     
                 
                   CYS GLU LYS ASN CYS SER CYS ASN SER CYS GLY CYS 
                 
                     
                 
                   HIS STOP; 
                 
                     
                 
                 
                 
               
                   [SEQ ID NO: 4] 
                     
                 
                 
                 
               
                   (2) MET ASP CYS CYS LYS ASN GLY CYS THR CYS ALA 
                     
                 
                     
                 
                   PRO ASN CYS LYS CYS ALA LYS ASP CYS LYS CYS; 
                 
                   and 
                 
             
                
               
            
             
                
                
                
                
                
                
                
                
                
                
               
            
             
                
               
            
             
                
                
                
                
               
            
           
         
         (3) sequences incorporating one or more conservative amino acid substitutions thereof. 
       
     
     
         3 . An isolated nucleic acid comprising DNA having at least 80% sequence identity to a DNA molecule having the sequence of nucleotide residues 1 to 66 of: 
       
         
           
                 
                 
               
                   [SEQ ID NO: 1] 
                     
                 
                 
                 
               
                   5′-ATG GAC TGC TGC AAG AAC GGT TGC ACC TGT GCC CCA 
                     
                 
                     
                 
                   AAT TGC AAA TGT GCC AAA GAC TGC AAA TGC TGC AAA 
                 
                     
                 
                   GGT TGT GAG TGC AAA AGC AAC CCA GAA TGC AAA TGT 
                 
                     
                 
                   GAG AAG AAC TGT TCA TGC AAC TCA TGT GGT TGT CAC 
                 
                     
                 
                   TGA-3′. 
                 
             
                
               
            
             
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         4 . The substantially purified metal binding protein of  claim 1  on a support. 
     
     
         5 . A vector including the nucleic acid sequence of  claim 3 . 
     
     
         6 . A metal binding composition comprising a substantially purified metal binding protein having an amino acid sequence analogous to a metal binding domain of a metal binding protein from a brine shrimp ( Artemia ). 
     
     
         7 . A method for reducing the concentration of a metal in a substrate, said method comprising the steps of:
 contacting a metal binding protein having an amino acid sequence analogous to at least one metal binding protein from a brine shrimp ( Artemia ) with said substrate to bind said metal to said metal binding protein; and   separating said bound metal from said substrate.   
     
     
         8 . The method of  claim 7  wherein said substrate is a fluid. 
     
     
         9 . The method of  claim 7  wherein said substrate is soil. 
     
     
         10 . The method of  claim 7  wherein said contacting step is under a high temperature condition. 
     
     
         11 . A method for the removal of metal from metal contaminated waste, said method comprising the steps of;
 contacting a metal binding protein having an amino acid sequence analogous to at least one metal binding protein sequence from a brine shrimp ( Artemia ) with said metal contaminated waste to bind said metal binding protein to said metal in said metal contaminated waste; and   separating said a bound metal from said contacted metal contaminated waste.   
     
     
         12 . The method of  claim 11  further comprising the additional step of:
 producing the metal binding protein in a modified organism.   
     
     
         13 . The method of  claim 7  wherein said metal binding protein is coupled to a support. 
     
     
         14 . The method of  claim 11  wherein said metal binding protein is coupled to a support. 
     
     
         15 . A modified organism producing a metal binding protein having an amino acid sequence substantially similar to SEQ ID NO: 2 and conservative amino acid substitutions thereof. 
     
     
         16 . The modified organism of  claim 15  wherein said modified organism is a transgenic plant. 
     
     
         17 . The modified organism of  claim 15  wherein said modified organism is a transgenic shrimp. 
     
     
         18 . The modified organism of  claim 15  wherein said modified organism is a bacteria. 
     
     
         19 . A method for producing at least one metal binding protein having an amino acid sequence analogous to at least one metal binding protein from a brine shrimp ( Artemia ) comprising the steps of:
 providing an expression system capable of producing at least one metal binding protein;   producing at least one metal binding protein having an amino acid sequence analogous to at least one metal binding protein from a brine shrimp ( Artemia ) utilizing said expression system; and   isolating said at least one metal binding protein having an amino acid sequence analogous to at least one metal binding protein from a brine shrimp ( Artemia ) from said expression system.