1oi8: Difference between revisions
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
[[Image:1oi8.jpg|left|200px]] | [[Image:1oi8.jpg|left|200px]] | ||
<!-- | |||
The line below this paragraph, containing "STRUCTURE_1oi8", 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_1oi8| PDB=1oi8 | SCENE= }} | ||
| | |||
}} | |||
'''5'-NUCLEOTIDASE (E. COLI) WITH AN ENGINEERED DISULFIDE BRIDGE (P90C, L424C)''' | '''5'-NUCLEOTIDASE (E. COLI) WITH AN ENGINEERED DISULFIDE BRIDGE (P90C, L424C)''' | ||
| Line 28: | Line 25: | ||
[[Category: Schultz-Heienbrok, R.]] | [[Category: Schultz-Heienbrok, R.]] | ||
[[Category: Straeter, N.]] | [[Category: Straeter, N.]] | ||
[[Category: | [[Category: Disulfide engineering,udp-sugar hydrolase]] | ||
[[Category: | [[Category: Domain movement]] | ||
[[Category: | [[Category: Hydrolase]] | ||
[[Category: | [[Category: Metalloprotein]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 03:52:45 2008'' | |||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | |||
Revision as of 03:52, 3 May 2008
5'-NUCLEOTIDASE (E. COLI) WITH AN ENGINEERED DISULFIDE BRIDGE (P90C, L424C)
OverviewOverview
Engineering disulfide bridges is a common technique to lock a protein movement in a defined conformational state. We have designed two double mutants of Escherichia coli 5'-nucleotidase to trap the enzyme in both an open (S228C, P513C) and a closed (P90C, L424C) conformation by the formation of disulfide bridges. The mutant proteins have been expressed, purified, and crystallized, to structurally characterize the designed variants. The S228C, P513C is a double mutant crystallized in two different crystal forms with three independent conformers, which differ from each other by a rotation of up to 12 degrees of the C-terminal domain with respect to the N-terminal domain. This finding, as well as an analysis of the domain motion in the crystal, indicates that the enzyme still exhibits considerable residual domain flexibility. In the double mutant that was designed to trap the enzyme in the closed conformation, the structure analysis reveals an unexpected intermediate conformation along the 96 degrees rotation trajectory between the open and closed enzyme forms. A comparison of the five independent conformers analyzed in this study shows that the domain movement of the variant enzymes is characterized by a sliding movement of the residues of the domain interface along the interface, which is in contrast to a classical closure motion where the residues of the domain interface move perpendicular to the interface.
About this StructureAbout this Structure
1OI8 is a Single protein structure of sequence from Escherichia coli. Full crystallographic information is available from OCA.
ReferenceReference
Trapping a 96 degrees domain rotation in two distinct conformations by engineered disulfide bridges., Schultz-Heienbrok R, Maier T, Strater N, Protein Sci. 2004 Jul;13(7):1811-22. PMID:15215524 Page seeded by OCA on Sat May 3 03:52:45 2008