Methods for determining tyrosine-DNA phosphodiesterase activity
AssigneeKelk Graduate of Applied Life Sciences
InventorsChen-Chen Kan, Ting-Jen Cheng
Life of this patent
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Abstract
The present invention provides human tyrosine-DNA phosphodiesterases (TDPs). In particular, the present invention provides novel recombinant nucleic acids and proteins, including mutant TDPs, vectors, and TDP-producing cells, as well as co-factors for enzyme activity. The present invention further provides methods for high through-put enzymatic assay systems utilizing the TDPs of the present invention.
Claims6
A method of determining tyrosine-DNA phosphodiesterase activity, comprising:
- a. providing:
- i. tyrosine-DNA phosphodiesterase:
- ii. p-nitrophenyl thymidine-3′-phosphate;
- and
- iii. manganese cation:
- b. combining said tyrosine-DNA phosphodiesterase, said p-nitrophenyl thymidine-3′-phosphate and said manganese cation under conditions wherein the tyrosine-DNA phosphodiesterase reacts with the p-nitrophenyl thymidine-3′-phosphate to release t,-nitrophenyl;
- and
- c. detecting said p-nitrophenyl, wherein said tyrosine-DNA phosphodiesterase is human tyrosine-DNA phosphodiesterase comprising amino acid sequence SEQ ID NQ:2.
The A method of determining tyrosine-DNA phosphodiesterase activity, comprising:
- a. providing:
- i. tyrosine-DNA phosphodiesterase:
- ii. p-nitrophenyl thymidine-3′-phosphate;
- and
- iii. manganese cation:
- b. combining said tyrosine-DNA phosphodiesterase, said p-nitrophenyl thymidine-3′-phosphate and said manganese cation under conditions wherein the tyrosine-DNA phosphodiesterase reacts with the p-nitrophenyl thymidine-3′-phosphate to release p-nitrophenyl;
- and
- c. detecting said p-nitrophenyl,
- wherein said tyrosine-DNA phosphodiesterase has an amino acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:8, 10, 12 and 14.
A method of determining tyrosine-DNA phosphodiesterase activity, comprising:
- a. providing:
- i. tyrosine-DNA phosphodiesterase;
- and
- ii. n-nitrophenyl thymidine-3′-phosphate;
- b. combining said tyrosine-DNA phosphodiesterase and said p-nitrophenyl thymidine-3′-phosphate under conditions wherein the tyrosine-DNA phosphodiesterase reacts with the n-nitrophenyl thymidine-3′-phosphate to release p-nitrophenyl;
- and
- c. detecting said p-nitrophenyl;
- wherein said tyrosine-DNA phosphodiesterase is human tyrosine-DNA phosphodiesterase comprising amino acid sequence SEQ ID NO:2.
A method of claim determining tyrosine-DNA phosphodiesterase activity, comprising:
- a. providing:
- i. a tyrosine-DNA phosphodiesterase;
- and
- ii. p-nitrophenyl thymidine-3′-phosphate;
- b. combining said tyrosine-DNA phosphodiesterase and said p-nitrophenyl thymidine-3′-phosphate under conditions wherein the tyrosine-DNA phosphodiesterase reacts with the n-nitrophenyl thymidine-3′-phosphate to release p-nitrophenyl;
- and
- c. detecting said p-nitrophenyl,
- wherein said tyrosine-DNA phosphodiesterase has an amino acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:8, 10, 12 and 14.
A method of determining modulation of tyrosine-DNA phosphodiesterase activity;
- comprising:
- a. providing:
- i. tyrosine-DNA phosphodiesterase;
- ii. a compound suspected of modulating tyrosine-DNA phosphodiesterase activity;
- and
- iii. p-nitrophenyl thymidine-3′-phosphate;
- b. combining said tyrosine-DNA phosphodiesterase and said p-nitrophenyl thymidine-3′-phosphate in the presence and absence of said compound suspected of modulating tyrosine-DNA phosphodiesterase activity under conditions wherein the tyrosine-DNA phosphodiesterase reacts with p-nitrophenyl thymidine-3′-phosphate to release p-nitrophenyl in the presence and absence of said compound suspected of modulating tyrosine-DNA phosphodiesterase activity;
- and
- c. detecting said p-nitrophenyl,
- wherein said tyrosine-DNA phosphodiesterase is human tyrosine-DNA phosphodiesterase comprising amino acid sequence SEQ ID NO:2.
A method of of determining modulation of tyrosine-DNA phosphodiesterase activity, comprising:
- a. providing;
- i. tyrosine-DNA phosphodiesterase;
- ii. a compound suspected of modulating tyrosine-DNA phosphodiesterase activity;
- and
- iii. p-nitrophenyl thymidine-3′-phosphate;
- b. combining said tyrosine-DNA phosphodiesterase and said p-nitrophenyl thymidine-3′-phosphate in the presence and absence of said compound suspected of modulating tyrosine-DNA phosphodiesterase activity under conditions wherein the tyrosine-DNA phosphodiesterase reacts with p-nitrophenyl thymidine-3′-phosphate to release p-nitrophenyl in the presence and absence of said compound suspected of modulating tyrosine-DNA phosphodiesterase activity;
- and
- c. detecting said p-nitrophenyl
- wherein said tyrosine-DNA phosphodiesterase has an amino acid sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NOS:8, 10, 12 and 14.
Dependencies are inferred from the claim text.