From Proteopedia
(Difference between revisions)
proteopedia linkproteopedia link
|
|
| Line 1: |
Line 1: |
| - | [[Image:1wuo.gif|left|200px]] | + | {{Seed}} |
| | + | [[Image:1wuo.png|left|200px]] |
| | | | |
| | <!-- | | <!-- |
| Line 9: |
Line 10: |
| | {{STRUCTURE_1wuo| PDB=1wuo | SCENE= }} | | {{STRUCTURE_1wuo| PDB=1wuo | SCENE= }} |
| | | | |
| - | '''Crystal structure of metallo-beta-lactamase IMP-1 mutant (D81A)'''
| + | ===Crystal structure of metallo-beta-lactamase IMP-1 mutant (D81A)=== |
| | | | |
| | | | |
| - | ==Overview==
| + | <!-- |
| - | Metallo-beta-lactamase IMP-1 is a di-Zn(II) metalloenzyme that efficiently hydrolyzes beta-lactam antibiotics. Wild-type (WT) IMP-1 has a conserved Asp-120(81) in the active site, which plays an important role in catalysis. To probe the catalytic role of Asp-120(81) in IMP-1, the IMP-1 mutants, D120(81)A and D120(81)E, were prepared by site-directed mutagenesis, and various kinetics studies were conducted. The IMP-1 mutants exhibited 10(2)-10(4)-fold drops in k(cat) values compared with WT despite the fact that they contained two Zn(II) ions in the active site. To evaluate the acid-base characteristics of Asp-120(81), the pH dependence for hydrolysis was examined by stopped-flow studies. No observable pK(a) values between pH 5 and 9 were found for WT and D120(81)A. The rapid mixing of equimolar amounts of nitrocefin and all enzymes failed to result in the detection of an anion intermediate of nitrocefin at 650 nm. These results suggest that Asp-120(81) of IMP-1 is not a factor in decreasing the pK(a) for the water bridging two Zn(II) ions and is not a proton donor to the anionic intermediate. In the case of D120(81)E, the nitrocefin hydrolysis product, which shows a maximum absorption at 460 nm, was bound to D120(81)E in the protonated form. The three-dimensional structures of D120(81)A and D120(81)E were also determined at 2.0 and 3.0 A resolutions, respectively. In the case of D120(81)E, the Zn-Zn distance was increased by 0.3 A compared with WT, due to the change in the coordination mode of Glu-120(81)OE1 and the positional shift in the conserved His-263(197) at the active site.
| + | The line below this paragraph, {{ABSTRACT_PUBMED_15788415}}, adds the Publication Abstract to the page |
| | + | (as it appears on PubMed at http://www.pubmed.gov), where 15788415 is the PubMed ID number. |
| | + | --> |
| | + | {{ABSTRACT_PUBMED_15788415}} |
| | | | |
| | ==About this Structure== | | ==About this Structure== |
| Line 29: |
Line 33: |
| | [[Category: Hydrolysis]] | | [[Category: Hydrolysis]] |
| | [[Category: Metallo-beta-lactamase]] | | [[Category: Metallo-beta-lactamase]] |
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 14:09:42 2008'' | + | |
| | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Tue Jul 29 03:03:57 2008'' |
Revision as of 00:04, 29 July 2008
Template:STRUCTURE 1wuo
Crystal structure of metallo-beta-lactamase IMP-1 mutant (D81A)
Template:ABSTRACT PUBMED 15788415
About this Structure
1WUO is a Single protein structure of sequence from Serratia marcescens. Full crystallographic information is available from OCA.
Reference
Probing the role of Asp-120(81) of metallo-beta-lactamase (IMP-1) by site-directed mutagenesis, kinetic studies, and X-ray crystallography., Yamaguchi Y, Kuroki T, Yasuzawa H, Higashi T, Jin W, Kawanami A, Yamagata Y, Arakawa Y, Goto M, Kurosaki H, J Biol Chem. 2005 May 27;280(21):20824-32. Epub 2005 Mar 23. PMID:15788415
Page seeded by OCA on Tue Jul 29 03:03:57 2008
