1fwd: Difference between revisions
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[[Image:1fwd.gif|left|200px]]<br /> | [[Image:1fwd.gif|left|200px]]<br /><applet load="1fwd" size="450" color="white" frame="true" align="right" spinBox="true" | ||
<applet load="1fwd" size="450" color="white" frame="true" align="right" spinBox="true" | |||
caption="1fwd, resolution 2.0Å" /> | caption="1fwd, resolution 2.0Å" /> | ||
'''KLEBSIELLA AEROGENES UREASE, C319A VARIANT AT PH 9.4'''<br /> | '''KLEBSIELLA AEROGENES UREASE, C319A VARIANT AT PH 9.4'''<br /> | ||
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==About this Structure== | ==About this Structure== | ||
1FWD is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Klebsiella_aerogenes Klebsiella aerogenes] with NI as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Urease Urease], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.1.5 3.5.1.5] | 1FWD is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Klebsiella_aerogenes Klebsiella aerogenes] with NI as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Urease Urease], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.1.5 3.5.1.5] Known structural/functional Sites: <scene name='pdbsite=ACT:Residue Implicated In Catalysis'>ACT</scene> and <scene name='pdbsite=NIL:Ni Metallocenter'>NIL</scene>. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1FWD OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: nickel metalloenzyme]] | [[Category: nickel metalloenzyme]] | ||
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Revision as of 16:08, 18 December 2007
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KLEBSIELLA AEROGENES UREASE, C319A VARIANT AT PH 9.4
OverviewOverview
Cys319 is located on a mobile flap covering the active site of Klebsiella, aerogenes urease but does not play an essential role in catalysis. Four, urease variants altered at position C319 range from having high activity, (C319A) to no measurable activity (C319Y), indicating Cys is not required, at this position, but its presence is highly influential [Martin, P. R., &, Hausinger, R. P. (1992) J. Biol. Chem. 267, 20024-20027]. Here, we present, 2.0 A resolution crystal structures of C319A, C319S, C319D, and C319Y, proteins and the C319A variant inhibited by acetohydroxamic acid. These, structures show changes in the hydration of the active site nickel ions, and in the position and flexibility of the active site flap. The C319Y, protein exhibits an alternate conformation of the flap, explaining its, lack of activity. The changes in hydration and conformation suggest that, there are suboptimal protein-solvent and protein-protein interactions in, the empty urease active site which contribute to urease catalysis., Specifically, we hypothesize that the suboptimal interactions may provide, a significant source of substrate binding energy, and such hidden energy, may be a common phenomenon for enzymes that contain mobile active site, loops and undergo an induced fit. The acetohydroxamic acid-bound structure, reveals a chelate interaction similar to those seen in other, metalloenzymes and in a small molecule nickel complex. The inhibitor, binding mode supports the proposed mode of urea binding. We complement, these structural studies with extended functional studies of C319A urease, to show that it has enhanced stability and resistance to inhibition by, buffers containing nickel ions. The near wild-type activity and enhanced, stability of the C319A variant make it useful for further studies of, urease structure-function relationships.
About this StructureAbout this Structure
1FWD is a Protein complex structure of sequences from Klebsiella aerogenes with NI as ligand. Active as Urease, with EC number 3.5.1.5 Known structural/functional Sites: and . Full crystallographic information is available from OCA.
ReferenceReference
Structures of Cys319 variants and acetohydroxamate-inhibited Klebsiella aerogenes urease., Pearson MA, Michel LO, Hausinger RP, Karplus PA, Biochemistry. 1997 Jul 1;36(26):8164-72. PMID:9201965
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