Haloalkyl derivatives of reporter molecules used to analyze metabolic activity in cells
US5576424A · kind A · utility
Assignee
Inventors
Key dates
| Filing date | Nov 8, 1994 |
| Grant date | Nov 19, 1996 |
| Priority date | — |
| Expiry date | Nov 8, 2014 |
Classification
- Technology area (CPC G)Physics
- CPC primaryG01N2333/924
- WIPO fieldMeasurement
- WIPO sectorInstruments
Abstract
The subject invention provides substrates useful for analyzing the metabolic activity in cells by improving the retention of a detectable reporter molecule only in intact cells where a particular enzyme is present. In particular, improved retention results from a two part process involving conjugation of haloalkyl-substituted derivatives of a reporter molecule with intracellular cysteine-containing peptides while unblocking the reporter molecule. The substrates have the form EQU XR-SPACER-REPORTER-BLOCK PA1 wherein -BLOCK is a group selected to be removable by action of a specific analyte, to give REPORTER spectral properties different from those of the substrate, PA1 -REPORTER- is a molecule that, when no longer bound to BLOCK by a BLOCK-REPORTER bond, has spectral properties different from those of the substrate, PA1 -SPACER- is a covalent linkage, and PA1 XR- is a haloalkyl moiety that can covalently react with an intracellular thiol (Z-S-H) to form a thioether conjugate (Z-S-R-). After the substrate enters the cells, the analyte removes BLOCK to make REPORTER detectable by the change in spectral properties, and the haloalkyl XR reacts with the intracellular thiol to form the th…
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.