Flea allantoinase proteins and uses thereof
View Patent ↗The present invention relates to flea allantoinase proteins; to flea allantoinase nucleic acid molecules, including those that encode such flea allantoinase proteins; to antibodies raised against such proteins; and to compounds that inhibit the activity of such proteins. The present invention also includes methods to obtain such proteins, nucleic acid molecules, antibodies, and inhibitory compounds. The present invention also includes therapeutic compositions comprising such inhibitory compounds, particularly those that specifically inhibit flea allantoinase activity, as well as the use of such therapeutic compositions to treat animals.
1. An isolated Ctenocephalides felis protein encoded by a nucleic acid molecule that hybridizes to a polynucleotide with a nucleic acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:8 and SEQ ID NO:11, under conditions comprising: (1) hybridizing in a solution comprising 1×SSC in the absence of helix destabilizing compounds, at a temperature of about 37° C.; and (2) washing in a solution comprising 1×SSC and in the absence of helix destabilizing compounds, at a temperature of about 47.5° C., wherein said isolated protein has allantoinase activity.
2. The protein of claim 1 , wherein said nucleic acid molecule is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:9 and fragments thereof, wherein said fragment comprises at least 25 contiguous nucleotides from a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:9 and encodes a protein having allantoinase activity.
3. The protein of claim 1 , wherein said protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:10, and fragments thereof, wherein said fragment has allantoinase activity.
4. A composition comprising an excipient and an isolated Ctenocephalides felis protein encoded by a nucleic acid molecule that hybridizes to a polynucleotide with a nucleic acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:8 and SEQ ID NO:11, under conditions comprising: (1) hybridizing in a solution comprising 1×SSC in the absence of helix destabilizing compounds, at a temperature of about 37° C.; and (2) washing in a solution comprising 1×SSC and in the absence of helix destabilizing compounds, at a temperature of about 47.5° C., wherein said isolated protein has allantoinase activity.
5. The composition of claim 4 , wherein said nucleic acid molecule is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:9 and fragments thereof, wherein said fragment comprises at least 25 contiguous nucleotides from a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:9 and encodes a protein having allantoinase activity.
6. The composition of claim 4 , wherein said protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:10, and fragments thereof, wherein said fragment has allantoinase activity.
7. The composition of claim 4 , wherein said composition further comprises a component selected from the group consisting of an adjuvant and a carrier.
8. A method to detect an inhibitor of flea allantoinase activity, said method comprising: (a) contacting an isolated protein of claim 1 with a putative inhibitory compound under conditions in which, in the absence of said compound, said protein has activity; and (b) determining if said putative inhibitory compound inhibits said activity.
9. The method of claim 8 , wherein said nucleic acid molecule is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:9 and fragments thereof, wherein said fragment comprises at least 25 contiguous nucleotides from a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:9 and encodes a protein having allantoinase activity.
10. The method of claim 8 , wherein said protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:10, and fragments thereof, wherein said fragment has allantoinase activity.
11. An isolated Ctenocephalides felis protein comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, and SEQ ID NO:10 and variants thereof that are at least 95% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, and SEQ ID NO:10, wherein said protein has allantoinase activity.
12. The protein of claim 11 , wherein said protein is encoded by a nucleic acid molecule selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:9 and fragments thereof, wherein said fragment comprises at least 25 contiguous nucleotides from a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:9 and encodes a protein having allantoinase activity.
13. The protein of claim 11 , wherein said protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, and SEQ ID NO:10.
14. A kit comprising the isolated protein of claim 1 .
15. The kit of claim 14 , wherein said nucleic acid molecule is selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:9 and fragments thereof, wherein said fragment comprises at least 25 contiguous nucleotides from a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:4, SEQ ID NO:6, and SEQ ID NO:9 and encodes a protein having allantoinase activity.
16. The kit of claim 14 , wherein said protein comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:7, SEQ ID NO:10, and fragments thereof, wherein said fragment has allantoinase activity.