Methods for identifying modulators of insulin receptor phosphorylation
AssigneeMax-Planck-Gessellschaft zur Forderung der Wissenschaften E.V.
InventorsAxel Ullrich, Edmund Hoppe, Niels Peter Hundahl M.o slashed.ller
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Abstract
The present invention relates to cell lines useful for the screening and identification of compounds that by modulating phosphotyrosine phosphatase activity, modulate insulin receptor type tyrosine kinase mediated signal transduction. Genetically engineered cells expressing IR in culture overcome the effect of insulin on morphology and adhesion when they are also coexpressing RPTP.alpha. or RPTP.epsilon.. Such engineered cell lines may be used to screen and identify non-toxic compounds that could elicit or modulate insulin signal transduction even in the absence of insulin.
Claims18
A method of determining whether a compound modulates phosphorylation of a phosphorylated insulin receptor in the presence of a receptor-type phosphotyrosine phosphatase, comprising:
- (a) contacting the compound with a mammalian cell in the presence of insulin, said cell cotransformed or cotransfected with DNA encoding insulin receptor and receptor-type phosphotyrosine phosphatase .alpha. or .epsilon. to coexpress insulin receptor and receptor-type phosphotyrosine phosphatase .alpha. or .epsilon.;
- and
- (b) comparing changes in the morphology of the cell in (a) with morphology of a control cell corresponding to the mammalian cell in (a) in the presence of insulin, but not exposed to the compound, wherein said change in morphology correlates with a change in the degree of phosphorylation of the insulin receptor,
- such that if the morphology of the cell in (a) differs from that of the control cell in (b), a compound that modulates the phosphorylation of the insulin receptor is identified.
A method of determining whether a compound increases phosphorylation of insulin receptor in the presence of a receptor-type phosphotyrosine phosphatase, comprising;
- (a) contacting the compound with a mammalian cell attached to a surface, said cell cotransformed or cotransfected with DNA encoding insulin receptor and receptor-type phosphotyrosine phosphatase .alpha. or .epsilon. to coexpress insulin receptor and receptor-type phosphotyrosine phosphatase .alpha. or .epsilon.;
- and
- (b) comparing changes in the morphology of the cell in (a) with morphology of a control cell not exposed to the compound, wherein said morphology comprises rounding up and detachment of the cell from the surface,
- such that if the morphology of the cell in (a) differs from the control cell in (b), a compound that increases the phosphorylation of the insulin receptor is identified.
Dependencies are inferred from the claim text.