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PDBsum entry 1y1b
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Hydrolase inhibitor
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PDB id
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1y1b
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References listed in PDB file
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Key reference
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Title
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Structural and functional study of an anemonia elastase inhibitor, A "nonclassical" kazal-Type inhibitor from anemonia sulcata.
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Authors
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H.Hemmi,
T.Kumazaki,
K.Yoshizawa-Kumagaye,
Y.Nishiuchi,
T.Yoshida,
T.Ohkubo,
Y.Kobayashi.
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Ref.
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Biochemistry, 2005,
44,
9626-9636.
[DOI no: ]
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PubMed id
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Abstract
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Anemonia elastase inhibitor (AEI) is a "nonclassical" Kazal-type elastase
inhibitor from Anemonia sulcata. Unlike many nonclassical inhibitors, AEI does
not have a cystine-stabilized alpha-helical (CSH) motif in the sequence. We
chemically synthesized AEI and determined its three-dimensional solution
structure by two-dimensional NMR spectroscopy. The resulting structure of AEI
was characterized by a central alpha-helix and a three-stranded antiparallel
beta-sheet of a typical Kazal-type inhibitor such as silver pheasant ovomucoid
third domain (OMSVP3), even though the first and fifth half-cystine residues
forming a disulfide bond in AEI are shifted both toward the C-terminus in
comparison with those of OMSVP3. Synthesized AEI exhibited unexpected strong
inhibition toward Streptomyces griseus protease B (SGPB). Our previous study
[Hemmi, H., et al. (2003) Biochemistry 42, 2524-2534] demonstrated that the
site-specific introduction of the engineered disulfide bond into the OMSVP3
molecule to form the CSH motif could produce an inhibitor with a narrower
specificity. Thus, the CSH motif-containing derivative of AEI (AEI analogue) was
chemically synthesized when a Cys(4)-Cys(34) bond was changed to a
Cys(6)-Cys(31) bond. The AEI analogue scarcely inhibited porcine pancreatic
elastase (PPE), even though it exhibited almost the same potent inhibitory
activity toward SGPB. For the molecular scaffold, essentially no structural
difference was detected between the two, but the N-terminal loop from Pro(5) to
Ile(7) near the putative reactive site (Met(10)-Gln(11)) in the analogue moved
by 3.7 A toward the central helix to form the introduced Cys(6)-Cys(31) bond.
Such a conformational change in the restricted region correlates with the
specificity change of the inhibitor.
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