Autotaxin: motility stimulating protein useful in cancer diagnosis and therapy

Abstract

The present invention relates, in general, to autotaxin. In particular, the present invention relates to a DNA segment encoding autotaxin; recombinant DNA molecules containing the DNA segment; cells containing the recombinant DNA molecule; a method of producing autotaxin; antibodies to autotaxin; and identification of functional domains in autotaxin.

1 . A DNA segment coding for a polypeptide comprising an amino acid sequence corresponding to autotaxin, or a fragment thereof having at least 5 amino acids.  
     
     
         2 . The DNA segment according to  claim 1 , wherein said DNA segment encodes the amino acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 11, SEQ ID NO: 26 through SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38 through SEQ ID NO: 52, SEQ ID NO: 66 and SEQ ID NO: 69.  
     
     
         3 . An isolated polypeptide comprising an amino acid sequence corresponding to autotaxin, or a fragment thereof having at least 5 amino acids.  
     
     
         4 . The polypeptide according to  claim 3 , wherein said amino acid sequence comprises the amino acid sequence selected from the group consisting of SEQ ID NO: 1 through SEQ ID NO: 11, SEQ ID NO: 26 through SEQ ID NO: 34, SEQ ID NO: 36, and SEQ ID NO: 38 through SEQ ID NO: 52, SEQ ID NO: 67 and SEQ ID NO: 69.  
     
     
         5 . An isolated polypeptide bound to a solid support, comprising an amino acid sequence corresponding to autotaxin, or a fragment thereof having at least 5 amino acids.  
     
     
         6 . The polypeptide according to  claim 5 , wherein said polypeptide comprises the amino acid sequence selected from the group consisting of the SEQ ID NO: 1 through SEQ ID NO: 11, SEQ ID NO: 26 through SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38 through SEQ ID NO: 52., SEQ ID NO: 67 and SEQ ID NO: 69.  
     
     
         7 . A recombinant DNA molecule comprising a vector and the DNA segment according to  claim 1 .  
     
     
         8 . A cell that contains the recombinant DNA molecule according to  claim 7 .  
     
     
         9 . An antibody having binding affinity for autotaxin, or binding fragment thereof.  
     
     
         10 . A method of producing a recombinant autotaxin polypeptide said method comprising: 
 culturing a cell containing the recombinant DNA molecule of  claim 7  under conditions such that the DNA segment is expressed, producing said polypeptide; and    isolating said polypeptide.    
     
     
         11 . A method of purifying the autotaxin peptide of  claim 3 , comprising the steps of: 
 i) collecting and concentrating supernatant from cultured A2058 human melanoma cells whereby a first preparation of said peptide is produced;    ii) salt fractionating said first preparation to produce a second peptide preparation;    iii) isolating said peptide from said second preparation so that said peptide is obtained in substantially pure form.    
     
     
         12 . The method of  claim 11 , wherein said isolating step is effected by column chromatography.  
     
     
         13 . An isolated DNA encoding an autotaxin protein or fragment thereof wherein said DNA includes a nucleic acid sequence selected from the group consisting of SEQ ID NO: 35, SEQ ID NO: 37 and SEQ ID NO: 38.  
     
     
         14 . The DNA segment according to  claim 1 , wherein said DNA fragment comprises any one of the SEQ ID NO: 12 through SEQ ID NO: 25, or SEQ ID NO: 39 through SEQ ID NO: 52.  
     
     
         15 . The DNA segment according to  claim 13  wherein said DNA segment comprises any one of the SEQ ID NO: 12 through SEQ ID NO: 25.  
     
     
         16 . An isolated polypeptide comprising an amino acid sequence corresponding to autotaxin.  
     
     
         17 . A polypeptide bound to a solid support and comprising an amino acid sequence corresponding to autotaxin.  
     
     
         18 . A recombinant autotaxin polypeptide according to  claim 3 .  
     
     
         19 . An isolated polypeptide according to claim 3 having cell motility activity.