Abstract: A functional linker for a polypeptide in which two alpha or beta globin-like domains are genetically fused is determined by screening a library of genetically fused polypeptides, in which the linker region is varied, for the ability to participate in the formation of hemoglobin-like protein, as measured by the protein's response to carbon monoxide. In a preferred embodiment, cells expressing the protein turn red as a result of carbon monoxide pressure.

Abstract: The present invention relates to a hemoglobin compositions stabilized against the formation of aggregates. The present invention further relates to methods of making such hemoglobin compositions.

Abstract: This invention relates to proteins with N-terminal modifications in their amino acid sequence that reduce or eliminate the methylation of the N-terminal methionine that occurs when such proteins are produced in bacteria such as E. coli.

Abstract: The present invention relates to modified hemoglobin-like compounds. The novel compounds include a globin-like polypeptide containing at least two di-alpha domains and multimeric hemoglobin-like proteins having a core hemoglobin-like moiety directly attached to at least two other hemoglobin-like moieties. The invention also relates to nucleic acid molecules encoding the novel polypeptides. Methods of making the multimeric hemoglobin-like proteins are also provided, as well as compositions containing the proteins.

Abstract: The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.

Abstract: The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.We further describe the production of tetrameric human hemoglobin and di-alpha/beta.sub.2 hemoglobin in the yeast Saccharomyces cerevisiae. The synthesis of the protein is directed by a synthetic promotor consisting of two functional parts, an upstream activator sequence (UAS) that confers inducible transcription by galactose from a consensus yeast transcriptional initiation site.

Abstract: Mutated protein having a sequence with a non-proline amino acid replacement for a proline at amino acid position 4. The mutated protein is not methylated on its N-terminus when expressed in bacteria. The mutated protein is a mutation of a methylated protein that is at least partially methylated on its N-terminus when expressed in bacteria.

Abstract: The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.

Abstract: The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.We further describe the production of tetrameric human hemoglobin and di-alpha/beta.sub.2 hemoglobin in the yeast Saccharomyces cerevisiae. The synthesis of the protein is directed by a synthetic promotor consisting of two functional parts, an upstream activator sequence (UAS) that confers inducible transcription by galactose from a consensus yeast transcriptional initiation site.

Abstract: Cysteine substitution mutants of alpha and/or beta globin mutants are produced by recombinant DNA techniques and used in the construction, intracellularly or otherwise, of mutant hemoglobins in which alpha- and beta-globin like subunits are crosslinked by disulfide bonds. Solutions of these mutant hemoglobins are used as blood substitutes. Preferably, these mutant hemoglobins contain further mutations which reduce their affinity for oxygen. Hemoglobins are preferably obtained by recombinant DNA techniques. Both alpha and beta globin chains can now be readily expressed, making possible the commercial production of wholly artificial hemoglobin, whether conventional or mutant in form. Solutions of wholly artificial hemoglobins are also used as blood substitutes. Expression of the alpha globin gene was substantially improved by means of a beta globin gene "header".

Abstract: A tracer, such as a radioisotope, is conjugated to a hemoglobin-like protein, such as normal hemoglobin, for use as an in vitro diagnostic imaging agent.

Abstract: The present invention relates to methods of lessening the severity and frequency of adverse effects associated with administration of extracellular hemoglobin comprising administration of a smooth muscle relaxant. Preferred smooth muscle relaxants are terbutaline and nifedipine, which can be administered in a prophylactic treatment, responsive regimented treatment or responsive intermittent treatment.

Abstract: The alpha subunits of hemoglobin, which in nature are formed as separate polypeptide chains which bind noncovalently to the beta subunits, are here provided in the form of the novel molecule di-alpha globin, a single polypeptide chain defined by connecting the two alpha subunits either directly via peptide bond or indirectly by a flexible amino acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Di-alpha globin is expressed by recombinant DNA techniques. Di-beta globin may be similarly obtained.

Abstract: DNA molecules which encode pseudodimeric globin-like polypeptides with an asymmetric cysteine mutation suitable for crosslinking two tetramers, or which encode pseudooligomeric globin-like polypeptides comprising four or more globin-like domains, are useful in the preparation of multimeric hemoglobin-like proteins.

Abstract: The present invention relates to regulators of cellular gene transcription, particularly inhibitors of cellular gene transactivating factors and in particular to inhibition of gene transcription in a viral host cell that is subject to regulation by proteins or factors that originate from a virus as well as conjugates or fusion products of the inhibitors and internalization molecules, pharmaceutical compositions that can be used to alleviate or prevent the manifestation of disease states that are the result of unregulated DNA transcription as a result of transactivation, methods of treating diseases that are caused or exacerbated by the presence of transactivating factors, and regulated gene therapy to achieve long term drug delivery of the inhibitors of the present invention. This invention can be applied both to cells with genetic abnormalities or to cells infected with a virus. Preferably, at least one protein of the protein-protein interactions is a transactivating factor.

Abstract: The invention relates to methods and compositions for delivering drugs to the body by binding them to hemoglobin. This technique stabilizes the drug and extends its half-life in the body. The drug may be chemically coupled to or adsorbed on the hemoglobin chain(s). Therapeutics are preferably linked to an artificial hemoglobin via a cysteine located deep within a cleft in the hemoglobin molecule.

Abstract: The present invention relates to a method for the production of a substantially protoporphyrin IX free hemoglobin solution comprising: rapidly heating a crude protoporphyrin IX-containing hemoglobin solution for a relatively short time and at a relatively high temperature to reduce protoporphyrin IX-containing hemoglobin to insignificant levels in said protoporphyrin IX-containing hemoglobin solution.

Abstract: The present invention provides blood substitutes comprised of recombinantly produced mutant hemoglobin having an osmolarity greater than 303 milliosmoles per liter and less than 800 milliosmoles per liter. Such hyperosmolar blood substitute additionally comprises a physiologically acceptable molecule less diffusible than dextrose.

Abstract: The present invention relates to a method for stimulating hematopoiesis in a mammal comprising administration of a therapeutically effective amount of a hemoglobin, including recombinant hemoglobin, and methods for treating cytopenias. These cytopenias include anemia, thrombocytopenia, lymphopenia, neutropenia and the like. The stimulation of hematopoiesis can occur both in vivo and ex vivo, as in the treatment of cytopenias associated with disease states, in cell culture or ex vivo expansion of bone marrow cells.

Abstract: Multimeric hemoglobin-like proteins are obtained by crosslinking cysteines of the component tetramers, or by genetically fusing globin-like domains of one tetramer with those of another, by means of an interdomain spacer sequence. Artificial cysteines are introduced selectively in a single globin-like domain per tetramer to control polymerization.