Abstract
Extracellular cysteine proteases, in particular cathepsin B, have been implicated in a variety of pathological processes. Selectively targeting labels of this enzyme are important tools to gain more detailed understanding of its specific roles. Starting from our recently developed irreversible epoxysuccinyl-based inhibitor (R-Gly-Gly-Leu-(2S,3S)-tEps-Leu-Pro-OH, R=OMe), we have synthesized two affinity labels, R=NH-(CH(2))(6)-NH-rhodamine B and R=NH-(CH(2))(6)-NH-biotin. Using MCF-7 cells, the labeled inhibitors were shown to be virtually non-cell-permeant. Moreover, affinity blot analysis with the biotinylated inhibitor allowed a highly sensitive and selective non-radioactive detection of active cathepsin B.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Affinity Labels / chemical synthesis*
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Affinity Labels / chemistry
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Biotin / analogs & derivatives*
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Biotin / chemical synthesis
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Biotin / chemistry
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Biotin / pharmacology
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Cathepsin B / chemistry*
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Cathepsin B / metabolism*
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Cathepsin L
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Cathepsins / chemistry
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Cathepsins / metabolism
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Cysteine Endopeptidases
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Endopeptidases*
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Humans
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Indicators and Reagents
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Kinetics
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Molecular Structure
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Oligopeptides / chemical synthesis*
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Oligopeptides / chemistry
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Oligopeptides / pharmacology
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Rhodamines / chemical synthesis*
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Rhodamines / chemistry
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Rhodamines / pharmacology
Substances
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Affinity Labels
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Indicators and Reagents
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Oligopeptides
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Rhodamines
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biotin-aminohexylamino-glycyl-glycyl-leucyl-epoxysuccinyl-leucyl-proline
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rhodamine B-aminohexylamino-glycyl-glycyl-leucyl-epoxysuccinyl-leucyl-proline
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Biotin
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Cathepsins
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Endopeptidases
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Cysteine Endopeptidases
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Cathepsin B
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CTSL protein, human
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Cathepsin L