Polypeptide cartilage-inducing factors found in bone

Abstract

Two proteins that are found is bone and that have in vivo chondrogenic/osteogenic activity in combination with a co-factor are described. Both proteins also were active in combination with EGF in the in vitro TGF-β assay. Each has a molecular weight of approximately 26,000 daltons by SDS-PAGE. Each is reduced to a single polypeptide indicating that the proteins are probably homodimers. One has an N-terminal sequence identical to that of human placenta-derived TGF-β whereas the other has an N-terminal sequence that is different from that of TGF-β derived from human placenta. The two proteins may be purified to homogeneity using RP-HPLC or acetic acid-urea gel electrophoresis.

We claim: 
     
       1. A composition for promoting proliferation of normal animal cells comprising: (a) a polypeptide that: (i) is active in the TGF-β assay; and   (ii) is a dimer having a molecular weight of approximately 26,000 daltons as determined by SDS-PAGE, whose chains each have the following N-terminal sequence: Ala-Leu-Asp-Ala-Ala-Tyr-Cys-Phe-Arq-Asn-Val-Gln-Asp-Asn-Cys-Cys-Leu-Arg-Pro-Leu-Tyr-Ile-Asp-Phe-Lys-Arg-Asp-Leu-Gly-Trp-;   a TGF-β activating agent; and   a pharmaceutically acceptable carrier, said polypeptide and activating agent being present in amounts that promote normal cell proliferation.     
     
     
       2. The composition of claim 1 wherein the activating agent is an EGF or a TGF-α. 
     
     
       3. A composition for promoting connective tissue deposition comprising: (a) an effective amount of a polypeptide that (i) is found in mammalian bone;   (ii) is active in the TGF-β assay;   (iii) is a co-factor for inducing cartilage formation in vivo;   (iv) is a dimer having an approximate molecular weight of 26,000 daltons as determined by SDS-PAGE; and   (v) is free of effective amounts of TGF-β activating agent or chondrogenic co-factor; and     (b) a pharmaceutically acceptable carrier.   
     
     
       4. A composition for promoting connective tissue depositon comprising: (a) an effective amount of a polypeptide that (i) is active in the TGF-β assay;   (ii) is a dimer having a molecular weight of approximately 26,000 daltons as determined by SDS-PAGE, whose chains each have the following N-terminal sequence: Ala-Leu-Asp-Ala-Ala-Tyr-Cys-Phe-Arg-Asn-Val-Gln-Asp-Asn-Cys-Cys-Leu-Arg-Pro-Leu-Tyr-Ile-Asp-Phe-Lys-Arg-Asp-Leu-Gly-Trp-; and   (iii) is free of effective amount of TGF-β activating agent or chondrogenic co-factor; and     (b) a pharmaceutically acceptable carrier. .Iadd.   
     
     
       5.  A composition for promoting proliferation of normal animal cells comprising: (a) a polypeptide that: (i) is active in the TGF-β assay; and   (ii) is a dimer having a molecular weight of approximately 26,000 daltons as determined by SDS-PAGE, whose chains each have the following N-terminal sequence: Ala-Leu-Asp-Ala-Ala-Tyr-Cys-Phe-Arq-Asn-Val-Gin-Asp-Asn-Cys-Cys-Leu-Arg-Pro-Leu-Tyr-Ile-Asp-Phe-Lys-Arg-Asp-Leu-Gly-Trp-; and     (b) a pharmaceutically acceptable carrier, said polypeptide being present in an amount that promotes normal cell proliferation. .Iaddend.