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PDBsum entry 3sic
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Complex(proteinase/inhibitor)
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PDB id
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3sic
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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Molecular recognition at the active site of subtilisin bpn': Crystallographic studies using genetically engineered proteinaceous inhibitor ssi (streptomyces subtilisin inhibitor).
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Authors
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Y.Takeuchi,
S.Noguchi,
Y.Satow,
S.Kojima,
I.Kumagai,
K.Miura,
K.T.Nakamura,
Y.Mitsui.
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Ref.
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Protein Eng, 1991,
4,
501-508.
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PubMed id
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Abstract
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Unlike trypsin-like serine proteases having only one conspicuous binding pocket
in the active site, subtilisin BPN' has two such pockets, the S1 and S4 pockets,
which accommodate the P1 and P4 residues of ligands (after Schechter and Berger
notation) respectively. Using computer graphics, the geometrical nature of the
two pockets was carefully examined and strategies for site-directed mutagenesis
studies were set up against a protein SSI (Streptomyces subtilisin inhibitor),
which is a strong proteinaceous inhibitor (or a substrate analogue) of
subtilisin BPN'. It was decided to convert the P1 residue, methionine 73, into
lysine (M73K) with or without additional conversion of the P4 residue,
methionine 70, into glycine (M70G). The crystal structures of the two complexes
of subtilisin BPN', one with the single mutant SSI (M73K) and the other with the
double mutant SSI (M73K, M70G) were solved showing that (i) small 'electrostatic
induced-fit movement' occurs in the S1 pocket upon introducing the terminal plus
charge of the lysine side chain, and (ii) large 'mechanical induced-fit
movement' occurs in the S4 pocket upon reducing the size of the P4 side chain
from methionine to glycine. In both (i) and (ii), the induced-fit movement
occurred in a concerted fashion involving both the enzyme and 'substrate' amino
acid residues. The term 'substrate-assisted stabilization' was coined to stress
the cooperative nature of the induced-fit movements.
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Secondary reference #1
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Title
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Refined crystal structure of the complex of subtilisin bpn' And streptomyces subtilisin inhibitor at 1.8 a resolution.
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Authors
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Y.Takeuchi,
Y.Satow,
K.T.Nakamura,
Y.Mitsui.
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Ref.
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J Mol Biol, 1991,
221,
309-325.
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PubMed id
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Secondary reference #2
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Title
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Crystal structure at 2.6 a resolution of the complex of subtilisin bpn' With streptomyces subtilisin inhibitor.
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Authors
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S.Hirono,
H.Akagawa,
Y.Mitsui,
Y.Iitaka.
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Ref.
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J Mol Biol, 1984,
178,
389-414.
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PubMed id
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Secondary reference #3
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Title
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Crystal structures of streptomyces subtilisin inhibitor and its complex with subtilisin bpn'.
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Authors
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Y.Mitsui,
Y.Satow,
Y.Watanabe,
S.Hirono,
Y.Iitaka.
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Ref.
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Nature, 1979,
277,
447-452.
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PubMed id
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