1dx9: Difference between revisions
No edit summary |
No edit summary |
||
Line 1: | Line 1: | ||
[[Image:1dx9.jpg|left|200px]] | [[Image:1dx9.jpg|left|200px]] | ||
<!-- | |||
The line below this paragraph, containing "STRUCTURE_1dx9", 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_1dx9| PDB=1dx9 | SCENE= }} | |||
}} | |||
'''W57A APOFLAVODOXIN FROM ANABAENA''' | '''W57A APOFLAVODOXIN FROM ANABAENA''' | ||
Line 27: | Line 24: | ||
[[Category: Romero, A.]] | [[Category: Romero, A.]] | ||
[[Category: Sancho, J.]] | [[Category: Sancho, J.]] | ||
[[Category: | [[Category: Flavoprotein]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 14:23:29 2008'' | |||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on |
Revision as of 14:23, 2 May 2008
W57A APOFLAVODOXIN FROM ANABAENA
OverviewOverview
Many flavoproteins are non-covalent complexes between FMN and an apoprotein. To understand better the stability of flavoproteins, we have studied the energetics of the complex between FMN and the apoflavodoxin from Anabaena PCC 7119 by a combination of site-directed mutagenesis, titration calorimetry, equilibrium binding constant determinations, and x-ray crystallography. Comparison of the strength of the wild type and mutant apoflavodoxin-FMN complexes and that of the complexes between wild type apoflavodoxin and shortened FMN analogues (riboflavin and lumiflavin) allows the dissection of the binding energy into contributions associated with the different parts of the FMN molecule. The estimated contribution of the phosphate is greatest, at 7 kcal mol(-1); that of the isoalloxazine is of around 5-6 kcal mol(-1) (mainly due to interaction with Trp-57 and Tyr-94 in the apoprotein) and the ribityl contributes least: around 1 kcal mol(-1). The stabilization of the complex is both enthalpic and entropic although the enthalpy contribution is dominant. Both the phosphate and the isoalloxazine significantly contribute to the enthalpy of binding. The ionic strength does not have a large effect on the stability of the FMN complex because, although it weakens the phosphate interactions, it strengthens the isoalloxazine-protein hydrophobic interactions. Phosphate up to 100 mM does not affect the strength of the riboflavin complex, which suggests the isoalloxazine and phosphate binding sites may be independent in terms of binding energy. Interestingly, we find crystallographic evidence of flexibility in one of the loops (57-62) involved in isoalloxazine binding.
About this StructureAbout this Structure
1DX9 is a Single protein structure of sequence from Anabaena sp.. Full crystallographic information is available from OCA.
ReferenceReference
Dissecting the energetics of the apoflavodoxin-FMN complex., Lostao A, El Harrous M, Daoudi F, Romero A, Parody-Morreale A, Sancho J, J Biol Chem. 2000 Mar 31;275(13):9518-26. PMID:10734100 Page seeded by OCA on Fri May 2 14:23:29 2008