Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Reissue Patent
2005-04-12
2005-04-12
Peselev, Elli (Department: 1623)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C536S004100, C536S018100, C549S356000, C549S382000
Reissue Patent
active
RE038723
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 formin-line-formulae description="In-line Formulae" end="lead"?XR-SPACER-REPORTER-BLOCKin-line-formulae description="In-line Formulae" end="tail"?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,-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,-SPACER- is a covalent linkage, andXR- 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 thioether conjugate inside the cells, which is well-retained in the cells.
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Haugland Richard P.
Mao Fei
Naleway John
Olson Nels
Sabnis Ram
Helfenstein Allegra
Molecular Probes, Inc.
Peselev Elli
Skaugset Anton
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