Nadph oxidase cytosolic cofactor mutant
AssigneeBoard of Regents, The University of Texas System
InventorsAnthony J. Valente, Robert A. Clark
Life of this patent
Estimating the term…
Abstract
A purified polypeptide includes an NADPH oxidase cytosolic cofactor polypeptide NOXA1 or p67phox having a C-terminal SH3 domain insertion mutation of at least three consecutive amino acids positioned between PEDL (SEQ ID NO: 35) and GIFPK (SEQ ID NO: 34) motifs of said SH3 domain so as to reduce activity of the purified polypeptide relative to the wild-type cofactor polypeptide. A process for treating, inhibiting, or prophylactically preventing a disease associated with the overproduction of reactive oxygen species involving NOXn where n is 1-4 inclusive includes the administration of a therapeutically effective amount with a purified polypeptide to a subject tissue that is overproducing reactive oxygen species through a NOXn pathway.
Claims11
A purified recombinantly expressed polypeptide comprising an NADPH oxidase cytosolic cofactor p67phox having a C-terminal SH3 domain insertion mutation of at least 3 consecutive amino acids positioned between Pro-Glu-Asp-Leu (SEQ ID NO:
- 35) and Gly-Ile-Phe-Pro-Lys (SEQ ID NO:
- 34) motifs of said SH3 domain so as to reduce activity of said cofactor relative to a wild-type cofactor of SEQ ID NO:
- 5 for a p67phox mutant, wherein said insertion mutation has a net hydropathic index of between −2.4 and 1.6;
- and an excision, addition or deletion mutation within bases 199-213, which is an activation domain of said cofactor to further reduce activity of said cofactor.
A purified recombinantly expressed polypeptide comprising an NADPH oxidase cytosolic cofactor p67phox having a C-terminal SH3 domain insertion mutation of at least 3 consecutive amino acids positioned between Pro-Glu-Asp-Leu (SEQ ID NO:
- 35) and Gly-Ile-Phe-Pro-Lys (SEQ ID NO:
- 34) motifs of said SH3 domain so as to reduce activity of said cofactor relative to a wild-type cofactor of SEQ ID NO:
- 5 for a p67phox mutant wherein said insertion mutation has a net hydropathic index of between −2.4 and 1.6;
- and a point mutation from tryptophan at residue 494 of said p67phox cofactor.
A purified recombinantly expressed polypeptide comprising an NADPH oxidase cytosolic cofactor NOXA1 (SEQ ID NO:
- 1) having an SH3 C-terminal domain insertion mutation of at least 3 consecutive amino acids inserted between Pro-Glu-Asp-Leu (SEQ ID NO:
- 35) and Gly-Ile-Phe-Pro-Lys (SEQ ID NO:
- 34) motifs of said SH3 C-terminal domain and at least one additional mutation selected from the group consisting of:
- an excision, addition or deletion mutation within bases 200-214 of an activation domain of said cofactor to further reduce activity of said cofactor, a point mutation changing a tryptophan at residue 436 of said NOXA1 cofactor, and a nonsilent mutation within a PB1 domain so as to reduce activity of said cofactor relative to a wild-type cofactor of SEQ ID NO:
- 1 for said NOXA1 cofactor, wherein said insertion mutation has a net hydropathic index of between −2.4 and 1.6.
Dependencies are inferred from the claim text.