Human sphingosine-1-phosphate phosphatase and inhibition methods
View Patent ↗The present invention provides polynucleotides and polypeptides of a human sphingosine-1-phosphate phosphatase, referred to herein as hSPP1. The polynucleotides and polypeptides are used to further provide expression vectors, host cells comprising the vectors, probes and primers, antibodies against the hSPP1 protein and polypeptides thereof, assays for the presence or expression of hSPP1 and assays for the identification of compounds that interact with hSPP1.
1. A method of determining whether a candidate compound is an inhibitor of a human sphingosine-1-phosphate phosphatase comprising:
(a) providing host cells harboring an expression vector that includes a polynucleotide selected from the group consisting of:
(i) a polynucleotide encoding a polypeptide having the amino acid sequence of SEQ ID NO: 2, and
(ii) a polynucleotide having the nucleotide sequence of SEQ ID NO: 1,
(b) contacting said cells with the candidate compound to permit the interaction of the candidate compound with the human sphingosine-1-phosphate phosphatase polypeptide, and
(c) determining whether the candidate compound is an inhibitor of said human sphingosine-1-phosphate phosphatase polypeptide by ascertaining the activity of the human sphingosine-1-phosphate phosphatase polypeptide in the presence of the candidate compound and comparing said activity to a measurement of the human sphingosine-1-phosphate phosphatase polypeptide activity of the cells before step (b).
2. A method of determining whether a candidate compound is an inhibitor of a human sphingosine- 1-phosphate phosphatase polypeptide comprising:
(a) providing a sample that includes a human sphingosine-1-phosphate phosphatase polypeptide having the amino acid sequence of SEQ ID NO: 2,
(b) contacting said sample with the candidate compound to permit the interaction of the candidate compound with the human sphingosine-1-phosphate phosphatase polypeptide, and
(c) determining whether the candidate compound is an inhibitor of said human sphingosine-1-phosphate phosphatase polypeptide by ascertaining the activity of the human sphingosine-1-phosphate phosphatase polypeptide in the presence of the candidate compound and comparing said activity to a measurement of the human sphingosine-1-phosphate phosphatase polypeptide activity of the cells before step (b).