Enzyme activity profiles
OwnerBenjamin F. Cravatt, Nadim Jessani +1inventors
InventorsBenjamin F. Cravatt, Nadim Jessani, Yongsheng Liu
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Abstract
Methods and compositions are provided for evaluating cellular status related to neoplasia. Affinity based probes, particularly having fluorophosphonates as a reactive functionality, are employed that react with a family of enzymes having a common catalytic activity, particularly serine/threonine hydrolases. The probe(s) are combined with the cell components and resulting conjugates are characterized, where the profile of reaction indicates cellular status. A novel serine/threonine hydrolase enzyme is provided.
Claims19
A method for evaluating cells in relation to their neoplastic state by determining the presence of a family of target enzymes having a common catalytic activity and having a common functionality in proximity to an active site associated with said common catalytic activity, said method comprising:
- combining the protein components of said cells with at least one affinity based probe characterized by preferentially binding to an active site of a family of enzymes and comprising a detectable entity, a reactive functionality for bonding to a protein functionality in spatial proximity to said active site, and a linker, each probe characterized by comprising the same reactive functionality group specific for said target enzymes, whereby said probes react with any target enzymes present in said cell;
- and
- determining the presence of the conjugate of each of said target enzymes with said probes by means of said detectable entity to obtain a profile of said target enzymes in said cell as said evaluation.
A method for evaluating cells in relation to their neoplastic state by determining the presence of a family of target serine/threonine hydrolases having a common functionality in proximity to an active site associated with a common catalytic activity, said method comprising:
- combining the protein components of said cells with at least one affinity based probe characterized by:
- preferentially binding to an active site of said family of hydrolases, comprising a detectable entity consisting of a ligand or fluorescer, a fluorophosphonyl group, and a linker, whereby said probes react with any target hydrolases present in said cell;
- and
- determining the presence of the conjugate of each of said target hydrolases with said probes by means of said detectable entity to obtain a profile of said target hydrolases in said cell as said evaluation.
A method for evaluating cells in relation to their neoplastic state by determining the presence of at least three of a family of target enzymes having a common catalytic activity and having a common functionality in proximity to an active site associated with said common catalytic activity, said method comprising:
- identifying the presence in said cells as compared to a control cell of at least three members of the group consisting of complement component 1s, PAF acetyl hydrolase isoform 1b, beta-subunit, FAAH, PPT-2, butyrylcholinesterase, p25, cathepsin A, PS_PL1, active uPA, esterase D, KIAA 1363, PAF-AH 2, p26, fatty acid synthase, APH, DPP8, lysophospholipase 1, alpha/beta hydrolase, peroxisomal long-chain acyl CoA thioesterase and angiotensinase C,
- wherein at least one of said three is FAAH, active uPA or KIAA 1363 whereby said neoplastic state is evaluated.
A composition comprising purified KIAA1363 characterized by being a serine/threonine hydrolase, present in melanoma and breast cancer cells, and comprising the amino acid sequence SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.