The SEP domain of p47 acts as a reversible competitive inhibitor of cathepsin L.
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.
Soukenik
Michael
M
Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle Str. 10, D-13125 Berlin, Germany.
Diehl
Anne
A
Leidert
Martina
M
Sievert
Volker
V
Büssow
Konrad
K
Leitner
Dietmar
D
Labudde
Dirk
D
Ball
Linda J
LJ
Lechner
Annette
A
Nägler
Dorit K
DK
Oschkinat
Hartmut
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002401:Cathepsin B / Q000737:chemistry / Q000378:metabolism; D056657:Cathepsin K; D056668:Cathepsin L; D002403:Cathepsins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology*; D017931:DNA Primers; D006801:Humans; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology*; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D015853:Cysteine Proteinase Inhibitors; D017931:DNA Primers; D010446:Peptide Fragments; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D002401:Cathepsin B; C534281:CTSL protein, human; D056668:Cathepsin L; C534130:CTSK protein, human; D056657:Cathepsin K