1yja
From Proteopedia
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| - | [[Image:1yja.jpg|left|200px]] | + | [[Image:1yja.jpg|left|200px]] |
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| - | '''SUBTILISIN BPN' 8397+1 (E.C. 3.4.21.14) (MUTANT WITH MET 50 REPLACED BY PHE, ASN 76 REPLACED BY ASP, GLY 169 REPLACED BY ALA, GLN 206 REPLACED BY CYS, ASN 218 REPLACED BY SER AND LYS 256 REPLACED BY TYR) (M50F, N76D, G169A, Q206C, N218S, AND K256Y) IN 20% DIMETHYLFORMAMIDE''' | + | {{Structure |
| + | |PDB= 1yja |SIZE=350|CAPTION= <scene name='initialview01'>1yja</scene>, resolution 1.8Å | ||
| + | |SITE= <scene name='pdbsite=CAT:Active+Site'>CAT</scene>, <scene name='pdbsite=STR:Occupied+By+A+Ca+Ion.+Strong+Binding+Site'>STR</scene> and <scene name='pdbsite=WEA:Occupied+By+A+Ca+Ion+And+A+Water+Molecule.+Weak+Binding+Site'>WEA</scene> | ||
| + | |LIGAND= <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene> and <scene name='pdbligand=HYD:HYDROXY GROUP'>HYD</scene> | ||
| + | |ACTIVITY= [http://en.wikipedia.org/wiki/Transferred_entry:_3.4.21.62 Transferred entry: 3.4.21.62], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.21.14 3.4.21.14] | ||
| + | |GENE= 8397+1 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1390 Bacillus amyloliquefaciens]) | ||
| + | }} | ||
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| + | '''SUBTILISIN BPN' 8397+1 (E.C. 3.4.21.14) (MUTANT WITH MET 50 REPLACED BY PHE, ASN 76 REPLACED BY ASP, GLY 169 REPLACED BY ALA, GLN 206 REPLACED BY CYS, ASN 218 REPLACED BY SER AND LYS 256 REPLACED BY TYR) (M50F, N76D, G169A, Q206C, N218S, AND K256Y) IN 20% DIMETHYLFORMAMIDE''' | ||
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==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 1YJA is a [ | + | 1YJA is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_amyloliquefaciens Bacillus amyloliquefaciens]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1YJA OCA]. |
==Reference== | ==Reference== | ||
| - | Breaking the low barrier hydrogen bond in a serine protease., Kidd RD, Sears P, Huang DH, Witte K, Wong CH, Farber GK, Protein Sci. 1999 Feb;8(2):410-7. PMID:[http:// | + | Breaking the low barrier hydrogen bond in a serine protease., Kidd RD, Sears P, Huang DH, Witte K, Wong CH, Farber GK, Protein Sci. 1999 Feb;8(2):410-7. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/10048334 10048334] |
[[Category: Bacillus amyloliquefaciens]] | [[Category: Bacillus amyloliquefaciens]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: zymogen]] | [[Category: zymogen]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 15:23:36 2008'' |
Revision as of 13:23, 20 March 2008
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| , resolution 1.8Å | |||||||
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| Sites: | , and | ||||||
| Ligands: | and | ||||||
| Gene: | 8397+1 (Bacillus amyloliquefaciens) | ||||||
| Activity: | Transferred entry: 3.4.21.62, with EC number 3.4.21.14 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
SUBTILISIN BPN' 8397+1 (E.C. 3.4.21.14) (MUTANT WITH MET 50 REPLACED BY PHE, ASN 76 REPLACED BY ASP, GLY 169 REPLACED BY ALA, GLN 206 REPLACED BY CYS, ASN 218 REPLACED BY SER AND LYS 256 REPLACED BY TYR) (M50F, N76D, G169A, Q206C, N218S, AND K256Y) IN 20% DIMETHYLFORMAMIDE
Overview
The serine protease subtilisin BPN' is a useful catalyst for peptide synthesis when dissolved in high concentrations of a water-miscible organic co-solvent such as N,N-dimethylformamide (DMF). However, in 50% DMF, the k(cat) for amide hydrolysis is two orders of magnitude lower than in aqueous solution. Surprisingly, the k(cat) for ester hydrolysis is unchanged in 50% DMF. To explain this alteration in activity, the structure of subtilisin 8397+1 was determined in 20, 35, and 50% (v/v) DMF to 1.8 A resolution. In 50% DMF, the imidazole ring of His64, the central residue of the catalytic triad, has rotated approximately 180 degrees around the Cbeta-Cgamma bond. Two new water molecules in the active site stabilize the rotated conformation. This rotation places His64 in an unfavorable geometry to interact with the other members of the catalytic triad, Ser221 and Asp32. NMR experiments confirm that the characteristic resonance due to the low barrier hydrogen bond between the His64 and Asp32 is absent in 50% DMF. These experiments provide a clear structural basis for the change in activity of serine proteases in organic co-solvents.
About this Structure
1YJA is a Single protein structure of sequence from Bacillus amyloliquefaciens. Full crystallographic information is available from OCA.
Reference
Breaking the low barrier hydrogen bond in a serine protease., Kidd RD, Sears P, Huang DH, Witte K, Wong CH, Farber GK, Protein Sci. 1999 Feb;8(2):410-7. PMID:10048334
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