1t2n: Difference between revisions

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[[Image:1t2n.jpg|left|200px]]<br /><applet load="1t2n" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:1t2n.jpg|left|200px]]<br /><applet load="1t2n" size="350" color="white" frame="true" align="right" spinBox="true"

caption="1t2n, resolution 1.80Å" />

'''Structure of a thermostable triple mutant of Bacillus subtilis lipase obtained through directed evolution'''<br />

==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.

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==

1T2N is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] with K as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [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] Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=1T2N OCA].

1T2N is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] with <scene name='pdbligand=K:'>K</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [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] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1T2N OCA].

==Reference==

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[[Category: alpha/beta hydrolase]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Wed Nov~~ 21 02:56:~~33 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:09:16 2008''

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@@ Line 1: / Line 1: @@
-[[Image:1t2n.jpg|left|200px]]<br /><applet load="1t2n" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1t2n.jpg|left|200px]]<br /><applet load="1t2n" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1t2n, resolution 1.80&Aring;" />
 '''Structure of a thermostable triple mutant of Bacillus subtilis lipase obtained through directed evolution'''<br />
 ==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.
+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==
-T2N is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] with K as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [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] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1T2N OCA].
+T2N is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] with <scene name='pdbligand=K:'>K</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [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] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1T2N OCA].
 ==Reference==
@@ Line 19: / Line 19: @@
 [[Category: alpha/beta hydrolase]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 02:56:33 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:09:16 2008''