Abstract: Disclosed is a method for the isolation and purification of polypeptides expressed in host cells by recombinant DNA techniques. A fused polypeptide is produced containing a desired polypeptide fused to additional amino acids. The additional amino acids define a leader sequence having properties exploitable in purification, a hinge region, and a cleavage site. The hinge region is cysteine-free and has a secondary structure which serves to expose the cleavage site to a selected endopeptidase. The method of the invention involves the production of a fused polypeptide which may be efficiently isolated by exploiting the properties of the leader sequence, and then efficiently cleaved at the cleavage site in an appropriate aqueous environment by virtue of the influence of the hinge on the cleavage agent/cleavage site reaction and other properties of the fused polypeptide.

Abstract: The invention disclosed herein provides methods and compositions for the computer-assisted design of morphogen analogs. Practice of the invention is enabled by the use of at least a portion of the atomic co-ordinates defining the three-dimensional structure of human transformation and differentiation factor-1 (hTDF-1) as a starting point in the design of the morphogen analogs. In addition, the invention provides methods for producing morphogen analogs of interest, and methods for testing whether the resulting analogs mimic or agonize human TDF-1-like biological activity. The invention also provides a family of morphogen analogs produced by such methods.

Abstract: The present invention relates to morphogen analogs, particularly analogs of a BMP, such as OP-1, that are agonists or antagonists of a BMP, such as OP-1, biological activity.

Abstract: Disclosed are a family of single-chain polypeptide constructs designed to agonize or mimic members of the TGF-&bgr; superfamily by binding to a cell surface receptor complementary to the superfamily member. The single-chain constructs of the invention called “morphons” contain in a single biologically active subunit interacting finger and heel regions which together define a tertiary protein structure complimentary to the ligand binding surface of a receptor that binds a TGF-&bgr; superfamily member. Also disclosed are truncated versions of the morphon constructs. Methods are disclosed for making and using single-chain morphons that have binding affinity for predetermined receptors of the TGF-&bgr; superfamily.

Abstract: Disclosed are a family of single-chain polypeptide constructs designed to agonize or mimic members of the TGF-&bgr; superfamily by binding to a cell surface receptor complementary to the superfamily member. The single-chain constructs of the invention called “morphons” contain in a single biologically active subunit interacting finger and heel regions which together define a tertiary protein structure complimentary to the ligand binding surface of a receptor that binds a TGF-&bgr; superfamily member . Also disclosed are truncated versions of the morphon constructs. Methods are disclosed for making and using single-chain morphons that have binding affinity for predetermined receptors of the TGF-&bgr; superfamily.

Abstract: The invention disclosed herein provides methods and compositions for the computer-assisted design of morphogen analogs. Practice of the invention is enabled by the use of at least a portion of the atomic co-ordinates defining the three-dimensional structure of human osteogenic protein-1 (hOP-1) as a starting point in the design of the morphogen analogs. In addition, the invention provides methods for producing morphogen analogs of interest, and methods for testing whether the resulting analogs mimic or agonize human OP-1-like biological activity. The invention also provides a family of morphogen analogs produced by such methods.

Abstract: The invention disclosed herein provides methods and compositions for the computer-assisted design of morphogen analogs. Practice of the invention is enabled by the use of at least a portion of the atomic co-ordinates defining the three-dimensional structure of human osteogenic protein-1 (hOP-1) as a starting point in the design of the morphogen analogs. In addition, the invention provides methods for producing morphogen analogs of interest, and methods for testing whether the resulting analogs mimic or agonize human OP-1-like biological activity. The invention also provides a family of morphogen analogs produced by such methods.

Abstract: Disclosed are a family of single-chain polypeptide constructs designed to agonize or mimic members of the TGF-.beta. superfamily by binding to a cell surface receptor complementary to the superfamily member. The single-chain constructs of the invention called "morphons" contain in a single biologically active subunit interacting finger and heel regions which together define a tertiary protein structure complimentary to the ligand binding surface of a receptor that binds a TGF-.beta. superfamily member. Also disclosed are truncated versions of the morphon constructs. Methods are disclosed for making and using single-chain morphons that have binding affinity for predetermined receptors of the TGF-.beta. superfamily.

Abstract: Protein analogues of tissue plasminogen activator (tPA) are described. The analogues exhibit the proteolytic function of natural tPA and optionally fibrin binding activity, but are molecules, generally of lower molecular weight than natural tPA, designed for efficient expression in prokaryotic host cell systems. The analogues can comprise a catalytic fragment of tPA or a catalytic fragment of tPA linked to a polypeptide which stabilizes the catalytic fragment, provides for efficient expression of the fragment or confers a fibrin binding capability. Fibrin binding polypeptides can be a polypeptide fragments derived from tPA which embody the fibrin binding domain(s) of natural tPA or they can be an exogenous (non-tPA) polypeptides of eukaryotic or prokaryotic origin which exhibit fibrin binding affinity such as the antigen binding fragment of an antifibrin immunoglobulin or the B domain of protein A. Genetic constructs for expression of the analogues are also provided.

Abstract: This invention pertains to a synthetic adhesive composition for use in aqueous environments. The composition comprises polypeptide chains having an .alpha.-helical structure in aqueous environments and capable of cohesive and adhesive interactions. The polypeptide chains comprise polar and apolar amino acids, the apolar and polar amino acids being arranged to define apolar and polar vertical spiraling stripes on the helix surface. The apolar stripes allow the polypeptide chains to aggregate into superhelical structures and the polar stripes allow interchain crosslinking within and between the superhelical structures.

Abstract: Disclosed is a novel polypeptide useful as a leader or trailer peptide moiety in recombinant DNA protein production techniques involving fused protein methodology. The polypeptide comprises an amphiphilic helix designed at the DNA level to have hydrophilic charged amino acid residues on one side of the barrel of the helix and nonpolar amino acid residues on the other side of the barrel of the helix. When DNA encoding the helix is attached to a gene encoding a protein of interest, high level expression is achieved and inclusion bodies are spontaneously formed. The inclusion bodies may be collected and purified easily by altering the ionic strength and/or pH of media used to dissolve the inclusion bodies. After purification, the fused protein is cleaved to separate the amphiphilic helix from the product.

Abstract: Disclosed is a novel polypeptide useful as a leader or trailer peptide moiety in recombinant DNA protein production techniques involving fused protein methodology. The polypeptide comprises an amphiphilic helix designed at the DNA level to have hydrophilic charged amino acid residues on one side of the barrel of the helix and nonpolar amino acid residues on the other side of the barrel of the helix. When DNA encoding the helix is attached to a gene encoding a protein of interest, high level expression is achieved and inclusion bodies are spontaneously formed. The inclusion bodies may be collected and purified easily by altering the ionic strength and/or pH of media used to dissolve the inclusion bodies. After purification, the fused protein is cleaved to separate the amphiphilic helix from the product.

Abstract: Disclosed is a novel polypeptide useful as a leader or trailer peptide moiety in recombinant DNA protein production techniques involving fused protein methodology The polypeptide comprises an amphiphilic helix designed at the DNA level to have hydrophilic charged amino acid residues on one side of the barrel of the helix and nonpolar amino acid residues on the other side of the barrel of the helix. When DNA encoding the helix is attached to a gene encoding a protein of interest, high level expression is achieved and inclusion bodies are spontaneously formed. The inclusion bodies may be collected and purified easily by altering the ionic strength and/or pH of media used to dissolve the inclusion bodies. After purification, the fused protein is cleaved to separate the amphiphilic helix from the product.

Abstract: This invention pertains to a synthetic adhesive composition for use in aqueous environments. The composition comprises polypeptide chains having an .alpha.-helical structure in aqueous environments and capable of cohesive and adhesive interactions. The polypeptide chains comprise polar and apolar amino acids, the apolar and polar amino acids being arranged to define apolar and polar vertical spiraling stripes on the helix surface. The apolar stripes allow the polypeptide chains to aggregate into superhelical structures and the polar stripes allow interchain crosslinking within and between the superhelical structures.

Abstract: Disclosed is a method for the isolation and purification of polypeptides expressed in host cells by recombinant DNA techniques. A fused polypeptide is produced containing a desired polypeptide fused to additional amino acids. The additional amino acids define a leader sequence having properties exploitable in purification, a hinge region, and a cleavage site. The hinge region is cysteine-free and has a secondary structure which serves to expose the cleavage site to a selected endopeptidase. The method of the invention involves the production of a fused polypeptide which may be efficiently isolated by exploiting the properties of the leader sequence, and then efficiently cleaved at the cleavage site in an appropriate aqueous environment by virtue of the influence of the hinge on the cleavage agent/cleavage site reaction and other properties of the fused polypeptide.