5sic: Difference between revisions

Line 1:

[[Image:5sic.gif|left|200px]]<br /><applet load="5sic" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:5sic.gif|left|200px]]<br /><applet load="5sic" size="350" color="white" frame="true" align="right" spinBox="true"

caption="5sic, resolution 2.2Å" />

'''MOLECULAR RECOGNITION AT THE ACTIVE SITE OF SUBTILISIN BPN': CRYSTALLOGRAPHIC STUDIES USING GENETICALLY ENGINEERED PROTEINACEOUS INHIBITOR SSI (STREPTOMYCES SUBTILISIN INHIBITOR)'''<br />

==Overview==

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.

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.

==About this Structure==

5SIC is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=5SIC OCA].

5SIC is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5SIC OCA].

==Reference==

Line 15:

[[Category: Subtilisin]]

[[Category: Mitsui, Y.]]

[[Category: Nakamura, K.T.]]

[[Category: Nakamura, K T.]]

[[Category: Takeuchi, Y.]]

[[Category: CA]]

[[Category: complex(proteinase/inhibitor)]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Tue Nov 20~~ 19:41:~~50 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 19:15:53 2008''

Navigation menu

Page actions

Page actions

Personal tools

Navigation

Search

Tools

@@ Line 1: / Line 1: @@
-[[Image:5sic.gif|left|200px]]<br /><applet load="5sic" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:5sic.gif|left|200px]]<br /><applet load="5sic" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="5sic, resolution 2.2&Aring;" />
 '''MOLECULAR RECOGNITION AT THE ACTIVE SITE OF SUBTILISIN BPN': CRYSTALLOGRAPHIC STUDIES USING GENETICALLY ENGINEERED PROTEINACEOUS INHIBITOR SSI (STREPTOMYCES SUBTILISIN INHIBITOR)'''<br />
 ==Overview==
-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.
+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.
 ==About this Structure==
-SIC is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=5SIC OCA].
+SIC is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Subtilisin Subtilisin], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.62 3.4.21.62] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5SIC OCA].
 ==Reference==
@@ Line 15: / Line 15: @@
 [[Category: Subtilisin]]
 [[Category: Mitsui, Y.]]
-[[Category: Nakamura, K.T.]]
+[[Category: Nakamura, K T.]]
 [[Category: Takeuchi, Y.]]
 [[Category: CA]]
 [[Category: complex(proteinase/inhibitor)]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 19:41:50 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 19:15:53 2008''