1t4m
From Proteopedia
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| - | [[Image:1t4m.jpg|left|200px]] | + | [[Image:1t4m.jpg|left|200px]] |
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| - | '''STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION''' | + | {{Structure |
| + | |PDB= 1t4m |SIZE=350|CAPTION= <scene name='initialview01'>1t4m</scene>, resolution 2.00Å | ||
| + | |SITE= | ||
| + | |LIGAND= <scene name='pdbligand=K:POTASSIUM ION'>K</scene> | ||
| + | |ACTIVITY= [http://en.wikipedia.org/wiki/Triacylglycerol_lipase Triacylglycerol lipase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.1.3 3.1.1.3] | ||
| + | |GENE= Lip A ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1423 Bacillus subtilis]) | ||
| + | }} | ||
| + | |||
| + | '''STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION''' | ||
| + | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 1T4M is a [ | + | 1T4M is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1T4M OCA]. |
==Reference== | ==Reference== | ||
| - | Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase., Acharya P, Rajakumara E, Sankaranarayanan R, Rao NM, J Mol Biol. 2004 Aug 27;341(5):1271-81. PMID:[http:// | + | Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase., Acharya P, Rajakumara E, Sankaranarayanan R, Rao NM, J Mol Biol. 2004 Aug 27;341(5):1271-81. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/15321721 15321721] |
[[Category: Bacillus subtilis]] | [[Category: Bacillus subtilis]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: alpha/beta hydrolase]] | [[Category: alpha/beta hydrolase]] | ||
| - | ''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 14:13:45 2008'' |
Revision as of 12:13, 20 March 2008
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| , resolution 2.00Å | |||||||
|---|---|---|---|---|---|---|---|
| Ligands: | |||||||
| Gene: | Lip A (Bacillus subtilis) | ||||||
| Activity: | Triacylglycerol lipase, with EC number 3.1.1.3 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION
Overview
Variation in gene sequences generated by directed evolution approaches often does not assure a minimalist design for obtaining a desired property in proteins. While screening for enhanced thermostability, structural information was utilized in selecting mutations that are generated by error-prone PCR. By this approach we have increased the half-life of denaturation by 300-fold compared to the wild-type Bacillus subtilis lipase through three point mutations generated by only two cycles of error-prone PCR. At lower temperatures the activity parameters of the thermostable mutants are unaltered. High-resolution crystal structures of the mutants show subtle changes, which include stacking of tyrosine residues, peptide plane flipping and a better anchoring of the terminus, that challenge rational design and explain the structural basis for enhanced thermostability. The approach may offer an efficient and minimalist solution for the enhancement of a desired property of a protein.
About this Structure
1T4M is a Single protein structure of sequence from Bacillus subtilis. Full crystallographic information is available from OCA.
Reference
Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase., Acharya P, Rajakumara E, Sankaranarayanan R, Rao NM, J Mol Biol. 2004 Aug 27;341(5):1271-81. PMID:15321721
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