1i2e: Difference between revisions
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
[[Image:1i2e.jpg|left|200px]] | [[Image:1i2e.jpg|left|200px]] | ||
<!-- | |||
The line below this paragraph, containing "STRUCTURE_1i2e", creates the "Structure Box" on the page. | |||
You may change the PDB parameter (which sets the PDB file loaded into the applet) | |||
or the SCENE parameter (which sets the initial scene displayed when the page is loaded), | |||
or leave the SCENE parameter empty for the default display. | |||
| | --> | ||
| | {{STRUCTURE_1i2e| PDB=1i2e | SCENE= }} | ||
}} | |||
'''Ribonuclease T1 V16A mutant, form I''' | '''Ribonuclease T1 V16A mutant, form I''' | ||
| Line 24: | Line 21: | ||
Hydrophobic core manipulations in ribonuclease T1., De Vos S, Backmann J, Prevost M, Steyaert J, Loris R, Biochemistry. 2001 Aug 28;40(34):10140-9. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/11513591 11513591] | Hydrophobic core manipulations in ribonuclease T1., De Vos S, Backmann J, Prevost M, Steyaert J, Loris R, Biochemistry. 2001 Aug 28;40(34):10140-9. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/11513591 11513591] | ||
[[Category: Aspergillus oryzae]] | [[Category: Aspergillus oryzae]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Backmann, J.]] | [[Category: Backmann, J.]] | ||
| Line 30: | Line 26: | ||
[[Category: Steyaert, J.]] | [[Category: Steyaert, J.]] | ||
[[Category: Vos, S De.]] | [[Category: Vos, S De.]] | ||
[[Category: | [[Category: Cavity creation]] | ||
[[Category: | [[Category: Hydrophobic core packing]] | ||
[[Category: | [[Category: Ribonuclease]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 19:29:29 2008'' | |||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | |||
Revision as of 19:29, 2 May 2008
Ribonuclease T1 V16A mutant, form I
OverviewOverview
Differential scanning calorimetry, urea denaturation, and X-ray crystallography were combined to study the structural and energetic consequences of refilling an engineered cavity in the hydrophobic core of RNase T1 with CH(3), SH, and OH groups. Three valines that cluster together in the major hydrophobic core of T1 were each replaced with Ala, Ser, Thr, and Cys. Compared to the wild-type protein, all these mutants reduce the thermodynamic stability of the enzyme considerably. The relative order of stability at all three positions is as follows: Val > Ala approximately equal to Thr > Ser. The effect of introducing a sulfhydryl group is more variable. Surprisingly, a Val --> Cys mutation in a hydrophobic environment can be as or even more destabilizing than a Val --> Ser mutation. Furthermore, our results reveal that the penalty for introducing an OH group into a hydrophobic cavity is roughly the same as the gain obtained from filling the cavity with a CH(3) group. The inverse equivalence of the behavior of hydroxyl and methyl groups seems to be crucial for the unique three-dimensional structure of the proteins. The importance of negative design elements in this context is highlighted.
About this StructureAbout this Structure
1I2E is a Single protein structure of sequence from Aspergillus oryzae. Full crystallographic information is available from OCA.
ReferenceReference
Hydrophobic core manipulations in ribonuclease T1., De Vos S, Backmann J, Prevost M, Steyaert J, Loris R, Biochemistry. 2001 Aug 28;40(34):10140-9. PMID:11513591 Page seeded by OCA on Fri May 2 19:29:29 2008