Nucleotide and protein sequence of mammastatin and methods of use

Abstract

A nucleic acid sequence encoding Mammastatin, a specific mammary cell growth inhibitor. Mammastatin is encoded by a single nucleic acid sequence and has an approximate molecular weight of 44 kDa in its inactive, non-phosphorylated form. Normal mammary cells express functional phosphorylated forms having approximate molecular weights of 53 kDa and 49 kDa. Metastatic mammary cells either do not express Mammastatin at all, or do not express the 53 kDa or 49 kDa, phosphorylated forms. Mammary cancer cells are inhibited in their growth by the administration of phosphorylated mammastatin.

1 .- 17 . (canceled) 
     
     
         18 . A method comprising the steps of:
 a) determining an amount of mammastatin in a patient's sample by contacting said mammastatin with an antibody that binds specifically to a mammastatin protein comprising SEQ ID NO: 1;   b) comparing the amount of determined mammastatin to a control, said control comprising an amount of mammastatin from a normal, non-breast cancer female; and   c) correlating an amount of mammastatin less than the control as a characteristic of breast cancer.   
     
     
         19 . The method of  claim 18 , wherein said antibody is a monoclonal antibody. 
     
     
         20 . The method of  claim 18 , wherein said antibody is selected from the group consisting of monoclonal antibody 7G6, monoclonal antibody 3C6, and monoclonal antibody 6B8. 
     
     
         21 . The method of  claim 18 , wherein said sample comprises mammary tissue, mammary cells, or fluids from said patient. 
     
     
         22 . The method of  claim 18 , wherein said patient's sample comprises blood or blood serum from said patient. 
     
     
         23 . The method of  claim 18 , where said step of determining an amount of mammastatin comprises use of an immunoassay. 
     
     
         24 . The method of  claim 23 , wherein the immunoassay is selected from the group consisting of a Western blot, a dot blot, and an enzyme-linked immunoabsorbent assay. 
     
     
         25 . The method of  claim 18 , wherein said control comprises an amount of mammastatin of at least 10 ng/ml. 
     
     
         26 . The method of  claim 18 , wherein said control comprises mammary cells, or mammary tissue, or fluid obtained from a normal, non-breast cancer female. 
     
     
         27 . A monoclonal antibody specific for a mammastatin protein comprising SEQ ID NO: 1. 
     
     
         28 . The monoclonal antibody of  claim 27 , wherein said mammastatin protein has an approximate molecular weight selected from the group consisting of 44 kD, 49 kD and 53 kD. 
     
     
         29 . The monoclonal antibody of  claim 27 , wherein said antibody is selected from the group consisting of monoclonal antibody 7G6, monoclonal antibody 3C6, and monoclonal antibody 6B8.