Retinoid metabolizing protein
Amino acid sequences and corresponding nucleic acid sequence of retinoid metabolizing protein found in human, mouse and zebrafish are described, as well as methods of using same.
1. A method of screening drugs for their effect on activity of a retinoid metabolizing protein, comprising exposing a cell that has been transfected with a nucleic acid molecule encoding the protein, or a descendent cell thereof, to a drug, wherein said transfected cell or descendent cell thereof expresses the protein, and determining the effect on the activity, wherein said protein oxidizes a retinoid, said nucleic acid molecule comprising a nucleotide sequence that hybridizes under high stringency conditions, wherein high stringency conditions include a wash step of about 0.2×SSC at 50° C., to a polynucleotide having the nucleotide sequence shown as SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:31.
2. A method of screening drugs for their effect on activity of a retinoid metabolizing protein, comprising exposing a cell that has been transfected with a nucleic acid molecule encoding the protein, or a descendent cell thereof, to a drug, wherein said transfected cell or descendent cell thereof expresses the protein, and determining the effect on the activity, wherein said protein oxidizes a retinoid, said nucleic acid molecule comprising a nucleotide sequence that encodes an amino acid sequence that is at least 90 percent identical to SEQ ID NO:4.
3. The method of claim 2 , wherein the nucleic acid molecule encodes an amino acid sequence that is at least 95 percent identical to SEQ ID NO:4.
4. The method of claim 2 , wherein the retinoid is a retinoic acid.
5. The method of claim 4 , wherein the retinoic acid is all-trans-retinoic acid.
6. A method of screening drugs for their effect on activity of a retinoid metabolizing protein comprising exposing a cell that has been transfected with a nucleic acid molecule encoding the protein, or a descendent cell thereof, to a drug, wherein said transfected cell or descendent cell thereof expresses the protein, and determining the effect on the activity, wherein said protein hydroxylates a retinoid at the C4-position of the β-ionone ring, said nucleic acid molecule comprising a nucleotide sequence that hybridizes under high stringency conditions, wherein high stringency conditions include a wash step of about 0.2×SSC at 50° C., to a nucleic acid molecule having the nucleotide sequence shown as SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:31.
7. A method of screening drugs for their effect on activity of a retinoid metabolizing protein comprising exposing a cell that has been transfected with a nucleic acid molecule encoding the protein, or a descendent cell thereof, to a drug, wherein said transfected cell or descendent cell thereof expresses the protein, and determining the effect on the activity, wherein said protein hydroxylates a retinoid at the C4-position of the β-ionone ring, said nucleic acid molecule comprising a nucleotide sequence that encodes an amino acid sequence that is at least 90 percent identical to SEQ ID NO:4.
8. The method of claim 7 , wherein the nucleic acid molecule encodes an amino acid sequence that is at least 95 percent identical to SEQ ID NO:4.
9. The method of claim 7 , wherein the retinoid is a retinoic acid.
10. The method of claim 9 , wherein the retinoic acid is all-trans-retinoic acid.
11. A method of screening drugs for their effect on activity of a retinoid metabolizing protein, comprising exposing a microsomal preparation comprising said protein to a drug and determining the effect on the activity, wherein the microsomal preparation comprises a recombinant protein expressed by a cell that has been transfected with a nucleic acid molecule encoding the protein, or by a descendent cell thereof, wherein said protein oxidizes a retinoid, said nucleic acid molecule comprising a nucleotide sequence that hybridizes under high stringency conditions, wherein high stringency conditions include a wash step of about 0.2×SSC at 50° C., to a polynucleotide having the nucleotide sequence shown as SEQ ID NO:3, SEQ ID NO::5, or SEQ ID NO:31.
12. A method of screening drugs for their effect on activity of a retinoid metabolizing protein, comprising exposing a microsomal preparation comprising said protein to a drug and determining the effect on the activity, wherein the microsomal preparation comprises a recombinant protein expressed by a cell that has been transfected with a nucleic acid molecule encoding the protein, or by a descendent cell thereof, wherein said protein oxidizes a retinoid, said nucleic acid molecule comprising a nucleotide sequence that encodes an amino acid sequence that is at least 90 percent identical to SEQ ID NO:4.
13. The method of claim 12 , wherein the nucleic acid molecule encodes an amino acid sequence that is at least 95 percent identical to SEQ ID NO:4.
14. The method of claim 12 , wherein the retinoid is a retinoic acid.
15. The method of claim 14 , wherein the retinoic acid is all-trans-retinoic acid.
16. A method of screening drugs for their effect on activity of a retinoid metabolizing protein comprising exposing a microsomal preparation comprising said protein to a drug and determining the effect on the activity, wherein the microsomal preparation comprises a recombinant protein expressed by a cell that has been transfected with a nucleic acid molecule encoding the protein, or by a descendent cell thereof, wherein said protein hydroxylates a retinoid at the C4-position of the β-ionone ring, said nucleic acid molecule comprising a nucleotide sequence that hybridizes under high stringency conditions, wherein high stringency conditions include a wash step of about 0.2×SSC at 50° C., to a nucleic acid molecule having the nucleotide sequence shown as SEQ ID NO:3, SEQ ID NO:5, or SEQ ID NO:31.
17. A method of screening drugs for their effect on activity of a retinoid metabolizing protein comprising exposing a microsomal preparation comprising said protein to a drug and determining the effect on the activity, wherein the microsomal preparation comprises a recombinant protein expressed by a cell that has been transfected with a nucleic acid molecule encoding the protein, or by a descendent cell thereof, wherein said protein hydroxylates a retinoid at the C4-position of the β-ionone ring, said nucleic acid molecule comprising a nucleotide sequence that encodes an amino acid sequence that is at least 90 percent identical to SEQ ID NO:4.
18. The method of claim 17 , wherein the nucleic acid molecule encodes an amino acid sequence that is at least 95 percent identical to SEQ ID NO:4.
19. The method of claim 17 , wherein the retinoid is a retinoic acid.
20. The method of claim 19 , wherein the retinoic acid is all-trans-retinoic acid.
21. The method of claim 1 , wherein the retinoid is a retinoic acid.
22. The method of claim 21 , wherein the retinoic acid is all-trans-retinoic acid.
23. The method of claim 6 , wherein the retinoid is a retinoic acid.
24. The method of claim 23 , wherein the retinoic acid is all-trans-retinoic acid.
25. The method of claim 11 , wherein the retinoid is a retinoic acid.
26. The method pf claim 25 , wherein the retinoic acid is all-trans-retinoic acid.
27. The method of claim 16 , wherein the retinoid is a retinoic acid.
28. The method of claim 27 , wherein the retinoic acid is all-trans-retinoic acid.
29. The method of claim 5 , wherein said protein oxidizes said all-trans retinoic acid at the C4-position of the β-ionone ring.
30. The method of claim 3 , wherein said protein oxidizes all-trans retinoic acid at the C4-position of the β-ionone ring.
31. The method of claim 15 , wherein said protein oxidizes said all-trans retinoic acid at the C4-position of the β-ionone ring.
32. The method of claim 8 , wherein said protein hydroxylates all-trans retinoic acid at the C4-position of the β-ionone ring.
33. The method of claim 22 , wherein said protein oxidizes said all-trans retinoic acid at the C4-position of the β-ionone ring.
34. The method of claim 13 , wherein said protein oxidizes all-trans retinoic acid at the C4-position of the β-ionone ring.
35. The method of claim 26 , wherein said protein oxidizes said all-trans retinoic acid at the C4-position of the β-ionone ring.
36. The method of claim 18 , wherein said protein hydroxylates all-trans retinoic acid at the C4-position of the β-ionone ring.