230l
From Proteopedia
(New page: 200px<br /><applet load="230l" size="450" color="white" frame="true" align="right" spinBox="true" caption="230l, resolution 1.9Å" /> '''T4 LYSOZYME MUTANT M6...) |
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| - | [[Image:230l.jpg|left|200px]]<br /><applet load="230l" size=" | + | [[Image:230l.jpg|left|200px]]<br /><applet load="230l" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="230l, resolution 1.9Å" /> | caption="230l, resolution 1.9Å" /> | ||
'''T4 LYSOZYME MUTANT M6L'''<br /> | '''T4 LYSOZYME MUTANT M6L'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The substitution of methionines with leucines within the interior of a | + | The substitution of methionines with leucines within the interior of a protein is expected to increase stability both because of a more favorable solvent transfer term as well as the reduced entropic cost of holding a leucine side chain in a defined position. Together, these two terms are expected to contribute about 1.4 kcal/mol to protein stability for each Met --> Leu substitution when fully buried. At the same time, this expected beneficial effect may be offset by steric factors due to differences in the shape of leucine and methionine. To investigate the interplay between these factors, all methionines in T4 lysozyme except at the amino-terminus were individually replaced with leucine. Of these mutants, M106L and M120L have stabilities 0.5 kcal/mol higher than wild-type T4 lysozyme, while M6L is significantly destabilized (-2.8 kcal/mol). M102L, described previously, is also destabilized (-0.9 kcal/mol). Based on this limited sample it appears that methionine-to-leucine substitutions can increase protein stability but only in a situation where the methionine side chain is fully or partially buried, yet allows the introduction of the leucine without concomitant steric interference. The variants, together with methionine-to-lysine substitutions at the same sites, follow the general pattern that substitutions at rigid, internal sites tend to be most destabilizing, whereas replacements at more solvent-exposed sites are better tolerated. |
==About this Structure== | ==About this Structure== | ||
| - | 230L is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacteriophage_t4 Bacteriophage t4] with CL and BME as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Lysozyme Lysozyme], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.17 3.2.1.17] Full crystallographic information is available from [http:// | + | 230L is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacteriophage_t4 Bacteriophage t4] with <scene name='pdbligand=CL:'>CL</scene> and <scene name='pdbligand=BME:'>BME</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Lysozyme Lysozyme], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.17 3.2.1.17] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=230L OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Lysozyme]] | [[Category: Lysozyme]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Baase, W | + | [[Category: Baase, W A.]] |
| - | [[Category: Drew, D | + | [[Category: Drew, D L.]] |
| - | [[Category: Eldridge, A | + | [[Category: Eldridge, A M.]] |
| - | [[Category: Gassner, N | + | [[Category: Gassner, N C.]] |
| - | [[Category: Lipscomb, L | + | [[Category: Lipscomb, L A.]] |
| - | [[Category: Matthews, B | + | [[Category: Matthews, B W.]] |
[[Category: Snow, S.]] | [[Category: Snow, S.]] | ||
[[Category: BME]] | [[Category: BME]] | ||
| Line 27: | Line 27: | ||
[[Category: o-glycosyl]] | [[Category: o-glycosyl]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:21:25 2008'' |
Revision as of 14:21, 21 February 2008
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T4 LYSOZYME MUTANT M6L
Overview
The substitution of methionines with leucines within the interior of a protein is expected to increase stability both because of a more favorable solvent transfer term as well as the reduced entropic cost of holding a leucine side chain in a defined position. Together, these two terms are expected to contribute about 1.4 kcal/mol to protein stability for each Met --> Leu substitution when fully buried. At the same time, this expected beneficial effect may be offset by steric factors due to differences in the shape of leucine and methionine. To investigate the interplay between these factors, all methionines in T4 lysozyme except at the amino-terminus were individually replaced with leucine. Of these mutants, M106L and M120L have stabilities 0.5 kcal/mol higher than wild-type T4 lysozyme, while M6L is significantly destabilized (-2.8 kcal/mol). M102L, described previously, is also destabilized (-0.9 kcal/mol). Based on this limited sample it appears that methionine-to-leucine substitutions can increase protein stability but only in a situation where the methionine side chain is fully or partially buried, yet allows the introduction of the leucine without concomitant steric interference. The variants, together with methionine-to-lysine substitutions at the same sites, follow the general pattern that substitutions at rigid, internal sites tend to be most destabilizing, whereas replacements at more solvent-exposed sites are better tolerated.
About this Structure
230L is a Single protein structure of sequence from Bacteriophage t4 with and as ligands. Active as Lysozyme, with EC number 3.2.1.17 Full crystallographic information is available from OCA.
Reference
Context-dependent protein stabilization by methionine-to-leucine substitution shown in T4 lysozyme., Lipscomb LA, Gassner NC, Snow SD, Eldridge AM, Baase WA, Drew DL, Matthews BW, Protein Sci. 1998 Mar;7(3):765-73. PMID:9541409
Page seeded by OCA on Thu Feb 21 16:21:25 2008
Categories: Bacteriophage t4 | Lysozyme | Single protein | Baase, W A. | Drew, D L. | Eldridge, A M. | Gassner, N C. | Lipscomb, L A. | Matthews, B W. | Snow, S. | BME | CL | Glycosidase | Hydrolase | O-glycosyl
