Abstract
The solution structure of the human p47 SEP domain in a construct comprising residues G1-S2-p47(171-270) was determined by NMR spectroscopy. A structure-derived hypothesis about the domains' function was formulated and pursued in binding experiments with cysteine proteases. The SEP domain was found to be a reversible competitive inhibitor of cathepsin L with a Ki of 1.5 microM. The binding of G1-S2-p47(171-270) to cathepsin L was mapped by biochemical assays and the binding interface was investigated by NMR chemical shift perturbation experiments.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Base Sequence
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Binding Sites
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Cathepsin B / chemistry
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Cathepsin B / metabolism
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Cathepsin K
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Cathepsin L
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Cathepsins / antagonists & inhibitors*
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Cathepsins / chemistry
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Cathepsins / metabolism
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Cysteine Endopeptidases
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Cysteine Proteinase Inhibitors / pharmacology*
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DNA Primers
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Humans
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Kinetics
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Magnetic Resonance Spectroscopy
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Models, Molecular
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Molecular Sequence Data
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Peptide Fragments / pharmacology*
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Protein Structure, Secondary
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Sequence Alignment
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Sequence Homology, Amino Acid
Substances
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Cysteine Proteinase Inhibitors
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DNA Primers
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Peptide Fragments
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Cathepsins
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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
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CTSK protein, human
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Cathepsin K