Analysis of protein kinases in live cells

An assay for detecting kinase activity comprising:

• contacting the one or more mutated kinases that comprise a mutation that enlarges an ATP binding pocket of the kinase with an ATP or ADP analog-nanoparticle conjugate capable of intracellular delivery of the ATP analog-nanoparticle conjugate, wherein the ATP or ADP analog comprises a detectable label;
• assaying the one or more mutated kinases under conditions in which the ATP or ADP analog-nanoparticle conjugate contacts the one or more mutated kinases, wherein the detectable label is transferred from the ATP analog-nanoparticle conjugate to a substrate of the one or more mutated kinases;
• and
• detecting the detectable label on the substrate, wherein the ATP or ADP analog only fluoresces upon contact with the ATP binding pocket of the kinase.

A system for detecting kinase activity, comprising:

• a well that comprises one or more mutated kinases, wherein the one or more mutated kinases comprise a mutation that enlarges an ATP binding pocket of the kinase,
• wherein the one or more mutated kinases are contacted with an ATP or ADP analog-nanoparticle conjugate capable of intracellular delivery of the ATP or ADP analog-nanoparticle conjugate,
• wherein the ATP or ADP analog comprises a detectable label, and after a predetermined period of time, detecting a detectable label on a substrate from the ATP or ADP analog-nanoparticle conjugate contacts the one or more mutated kinases, and
• wherein the detectable label was transferred from the ATP or ADP analog-nanoparticle conjugate to the substrate of the one or more mutated kinases, and
• wherein the ATP or ADP analog only fluoresces upon contact with the ATP binding pocket of the kinase.