1ft7

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(New page: 200px<br /><applet load="1ft7" size="450" color="white" frame="true" align="right" spinBox="true" caption="1ft7, resolution 2.2&Aring;" /> '''AAP COMPLEXED WITH L-...)
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[[Image:1ft7.jpg|left|200px]]<br /><applet load="1ft7" size="450" color="white" frame="true" align="right" spinBox="true"
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[[Image:1ft7.jpg|left|200px]]<br /><applet load="1ft7" size="350" color="white" frame="true" align="right" spinBox="true"
caption="1ft7, resolution 2.2&Aring;" />
caption="1ft7, resolution 2.2&Aring;" />
'''AAP COMPLEXED WITH L-LEUCINEPHOSPHONIC ACID'''<br />
'''AAP COMPLEXED WITH L-LEUCINEPHOSPHONIC ACID'''<br />
==Overview==
==Overview==
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The nature of the interaction of the transition-state analogue inhibitor, L-leucinephosphonic acid (LPA) with the leucine aminopeptidase from, Aeromonas proteolytica (AAP) was investigated. LPA was shown to be a, competitive inhibitor at pH 8.0 with a K(i) of 6.6 microM. Electronic, absorption spectra, recorded at pH 7.5 of [CoCo(AAP)], [CoZn(AAP)], and, [ZnCo(AAP)] upon addition of LPA suggest that LPA interacts with both, metal ions in the dinuclear active site. EPR studies on the, Co(II)-substituted forms of AAP revealed that the environments of the, Co(II) ions in both [CoZn(AAP)] and [ZnCo(AAP)] become highly asymmetric, and constrained upon the addition of LPA and clearly indicate that LPA, interacts with both metal ions. The X-ray crystal structure of AAP, complexed with LPA was determined at 2.1 A resolution. The X-ray, crystallographic data indicate that LPA interacts with both metal centers, in the dinuclear active site of AAP and a single oxygen atom bridge is, absent. Thus, LPA binds to the dinuclear active site of AAP as an, eta-1,2-mu-phosphonate with one ligand to the second metal ion provided by, the N-terminal amine. A structural comparison of the binding of, phosphonate-containing transition-state analogues to the mono- and, bimetallic peptidases provides insight into the requirement for the second, metal ion in bridged bimetallic peptidases. On the basis of the results, obtained from the spectroscopic and X-ray crystallographic data presented, herein along with previously reported mechanistic data for AAP, a new, catalytic mechanism for the hydrolysis reaction catalyzed by AAP is, proposed.
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The nature of the interaction of the transition-state analogue inhibitor L-leucinephosphonic acid (LPA) with the leucine aminopeptidase from Aeromonas proteolytica (AAP) was investigated. LPA was shown to be a competitive inhibitor at pH 8.0 with a K(i) of 6.6 microM. Electronic absorption spectra, recorded at pH 7.5 of [CoCo(AAP)], [CoZn(AAP)], and [ZnCo(AAP)] upon addition of LPA suggest that LPA interacts with both metal ions in the dinuclear active site. EPR studies on the Co(II)-substituted forms of AAP revealed that the environments of the Co(II) ions in both [CoZn(AAP)] and [ZnCo(AAP)] become highly asymmetric and constrained upon the addition of LPA and clearly indicate that LPA interacts with both metal ions. The X-ray crystal structure of AAP complexed with LPA was determined at 2.1 A resolution. The X-ray crystallographic data indicate that LPA interacts with both metal centers in the dinuclear active site of AAP and a single oxygen atom bridge is absent. Thus, LPA binds to the dinuclear active site of AAP as an eta-1,2-mu-phosphonate with one ligand to the second metal ion provided by the N-terminal amine. A structural comparison of the binding of phosphonate-containing transition-state analogues to the mono- and bimetallic peptidases provides insight into the requirement for the second metal ion in bridged bimetallic peptidases. On the basis of the results obtained from the spectroscopic and X-ray crystallographic data presented herein along with previously reported mechanistic data for AAP, a new catalytic mechanism for the hydrolysis reaction catalyzed by AAP is proposed.
==About this Structure==
==About this Structure==
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1FT7 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Vibrio_proteolyticus Vibrio proteolyticus] with ZN, K and PLU as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Bacterial_leucyl_aminopeptidase Bacterial leucyl aminopeptidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.11.10 3.4.11.10] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1FT7 OCA].
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1FT7 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Vibrio_proteolyticus Vibrio proteolyticus] with <scene name='pdbligand=ZN:'>ZN</scene>, <scene name='pdbligand=K:'>K</scene> and <scene name='pdbligand=PLU:'>PLU</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Bacterial_leucyl_aminopeptidase Bacterial leucyl aminopeptidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.11.10 3.4.11.10] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1FT7 OCA].
==Reference==
==Reference==
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[[Category: zinc]]
[[Category: zinc]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 15:15:25 2007''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:42:25 2008''

Revision as of 10:42, 21 February 2008

Image:1ft7.jpg

1ft7, resolution 2.2Å

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AAP COMPLEXED WITH L-LEUCINEPHOSPHONIC ACID

Overview

The nature of the interaction of the transition-state analogue inhibitor L-leucinephosphonic acid (LPA) with the leucine aminopeptidase from Aeromonas proteolytica (AAP) was investigated. LPA was shown to be a competitive inhibitor at pH 8.0 with a K(i) of 6.6 microM. Electronic absorption spectra, recorded at pH 7.5 of [CoCo(AAP)], [CoZn(AAP)], and [ZnCo(AAP)] upon addition of LPA suggest that LPA interacts with both metal ions in the dinuclear active site. EPR studies on the Co(II)-substituted forms of AAP revealed that the environments of the Co(II) ions in both [CoZn(AAP)] and [ZnCo(AAP)] become highly asymmetric and constrained upon the addition of LPA and clearly indicate that LPA interacts with both metal ions. The X-ray crystal structure of AAP complexed with LPA was determined at 2.1 A resolution. The X-ray crystallographic data indicate that LPA interacts with both metal centers in the dinuclear active site of AAP and a single oxygen atom bridge is absent. Thus, LPA binds to the dinuclear active site of AAP as an eta-1,2-mu-phosphonate with one ligand to the second metal ion provided by the N-terminal amine. A structural comparison of the binding of phosphonate-containing transition-state analogues to the mono- and bimetallic peptidases provides insight into the requirement for the second metal ion in bridged bimetallic peptidases. On the basis of the results obtained from the spectroscopic and X-ray crystallographic data presented herein along with previously reported mechanistic data for AAP, a new catalytic mechanism for the hydrolysis reaction catalyzed by AAP is proposed.

About this Structure

1FT7 is a Single protein structure of sequence from Vibrio proteolyticus with , and as ligands. Active as Bacterial leucyl aminopeptidase, with EC number 3.4.11.10 Full crystallographic information is available from OCA.

Reference

Inhibition of the aminopeptidase from Aeromonas proteolytica by L-leucinephosphonic acid. Spectroscopic and crystallographic characterization of the transition state of peptide hydrolysis., Stamper C, Bennett B, Edwards T, Holz RC, Ringe D, Petsko G, Biochemistry. 2001 Jun 19;40(24):7035-46. PMID:11401547

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