Production and use of multimeric hemoglobins
First Claim
1. A non-naturally occurring pseudotetrameric hemoglobin-like protein comprising a pseudodimeric polypeptide having two substantially homologous globin-like domains, one of which is mutated to provide an asymmetric crosslinkable cysteine residue, wherein the corresponding residue in the other globin-like domain of said pseudodimeric polypeptide is an amino acid other than cysteine, and wherein said pseudotetramer further comprises at least one other globin-like domain-bearing polypeptide, and wherein the asymmetric crosslinkable cysteine residue is a mutation in the alpha globin-like domain corresponding to a human alpha globin position selected from the group consisting of lys7, thr8, lys11, ala12, ,gly15, lys16, gly18, ala19, his20, glu23, and thr24.
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Accused Products
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.
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Citations
7 Claims
- 1. A non-naturally occurring pseudotetrameric hemoglobin-like protein comprising a pseudodimeric polypeptide having two substantially homologous globin-like domains, one of which is mutated to provide an asymmetric crosslinkable cysteine residue, wherein the corresponding residue in the other globin-like domain of said pseudodimeric polypeptide is an amino acid other than cysteine, and wherein said pseudotetramer further comprises at least one other globin-like domain-bearing polypeptide, and wherein the asymmetric crosslinkable cysteine residue is a mutation in the alpha globin-like domain corresponding to a human alpha globin position selected from the group consisting of lys7, thr8, lys11, ala12, ,gly15, lys16, gly18, ala19, his20, glu23, and thr24.
- 3. A non-naturally occurring pseudotetrameric hemoglobin-like protein comprising a pseudodimeric polypeptide having two substantially homologous globin-like domains, one of which is mutated to provide an asymmetric crosslinkable cysteine residue, wherein the corresponding residue in the other globin-like domain of said pseudodimeric polypeptide is an amino acid other than cysteine, and wherein said pseudotetramer further comprises at least one other globin-like domain-bearing polypeptide, and wherein the asymmetric crosslinkable cystine residue is a mutation in the beta globin-like domain corresponding to a human beta globin position selected from the group consisting of his2, leu3, thr4, glu6, ser9, thr12, ala13, gly16, lys17, val18, asn19, val20, asp21, and glu22.
- 6. A recombinant mutant hemoglobin-like protein comprising a mutation in a beta-globin like domain at amino acid position 93, wherein the cysteine at position 93 is replaced with an amino acid chosen from the group consisting of alanine and threonine.
Specification