1ggv: Difference between revisions

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CRYSTAL STRUCTURE OF THE C123S MUTANT OF DIENELACTONE HYDROLASE (DLH) BOUND WITH THE PMS MOIETY OF THE PROTEASE INHIBITOR, PHENYLMETHYLSULFONYL FLUORIDE (PMSF)

OverviewOverview

The structure of DLH (C123S) with PMS bound was solved to 2.5 A resolution (R factor = 15.1%). PMSF in 2-propanol was delivered directly to crystals in drops and unexpectedly caused the crystals to dissolve. New crystals displaying a different morphology emerged within 2 h in situ, a phenomenon that appears to be described for the first time. The changed crystal form reflected altered crystal-packing arrangements elicited by structural changes to the DLH (C123S) molecule on binding inhibitor. The new unit cell remained in the P2(1)2(1)2(1) space group but possessed different dimensions. The structure showed that PMS binding in DLH (C123S) caused conformational changes in the active site and in four regions of the polypeptide chain that contain reverse turns. In the active site, residues with aromatic side chains were repositioned in an edge-to-face cluster around the PMS phenyl ring. Their redistribution prevented restabilization of the triad His202 side chain, which was disordered in electron-density maps. Movements of other residues in the active site were shown to be related to the four displaced regions of the polypeptide chain. Their implied synergy suggests that DLH may be able to accommodate and catalyse a range of compounds unrelated to the natural substrate owing to an inherent coordinated flexibility in its overall structure. Implications for mechanism and further engineering studies are discussed.

About this StructureAbout this Structure

1GGV is a Single protein structure of sequence from Pseudomonas putida. Active as Carboxymethylenebutenolidase, with EC number 3.1.1.45 Full crystallographic information is available from OCA.

ReferenceReference

Structure of the C123S mutant of dienelactone hydrolase (DLH) bound with the PMS moiety of the protease inhibitor phenylmethylsulfonyl fluoride (PMSF)., Robinson A, Edwards KJ, Carr PD, Barton JD, Ewart GD, Ollis DL, Acta Crystallogr D Biol Crystallogr. 2000 Nov;56(Pt 11):1376-84. PMID:11053834

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-[[Image:1ggv.gif|left|200px]]<br /><applet load="1ggv" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1ggv.gif|left|200px]]<br /><applet load="1ggv" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1ggv, resolution 2.50&Aring;" />
 '''CRYSTAL STRUCTURE OF THE C123S MUTANT OF DIENELACTONE HYDROLASE (DLH) BOUND WITH THE PMS MOIETY OF THE PROTEASE INHIBITOR, PHENYLMETHYLSULFONYL FLUORIDE (PMSF)'''<br />
 ==Overview==
-The structure of DLH (C123S) with PMS bound was solved to 2.5 A resolution, (R factor = 15.1%). PMSF in 2-propanol was delivered directly to crystals, in drops and unexpectedly caused the crystals to dissolve. New crystals, displaying a different morphology emerged within 2 h in situ, a phenomenon, that appears to be described for the first time. The changed crystal form, reflected altered crystal-packing arrangements elicited by structural, changes to the DLH (C123S) molecule on binding inhibitor. The new unit, cell remained in the P2(1)2(1)2(1) space group but possessed different, dimensions. The structure showed that PMS binding in DLH (C123S) caused, conformational changes in the active site and in four regions of the, polypeptide chain that contain reverse turns. In the active site, residues, with aromatic side chains were repositioned in an edge-to-face cluster, around the PMS phenyl ring. Their redistribution prevented restabilization, of the triad His202 side chain, which was disordered in electron-density, maps. Movements of other residues in the active site were shown to be, related to the four displaced regions of the polypeptide chain. Their, implied synergy suggests that DLH may be able to accommodate and catalyse, a range of compounds unrelated to the natural substrate owing to an, inherent coordinated flexibility in its overall structure. Implications, for mechanism and further engineering studies are discussed.
+The structure of DLH (C123S) with PMS bound was solved to 2.5 A resolution (R factor = 15.1%). PMSF in 2-propanol was delivered directly to crystals in drops and unexpectedly caused the crystals to dissolve. New crystals displaying a different morphology emerged within 2 h in situ, a phenomenon that appears to be described for the first time. The changed crystal form reflected altered crystal-packing arrangements elicited by structural changes to the DLH (C123S) molecule on binding inhibitor. The new unit cell remained in the P2(1)2(1)2(1) space group but possessed different dimensions. The structure showed that PMS binding in DLH (C123S) caused conformational changes in the active site and in four regions of the polypeptide chain that contain reverse turns. In the active site, residues with aromatic side chains were repositioned in an edge-to-face cluster around the PMS phenyl ring. Their redistribution prevented restabilization of the triad His202 side chain, which was disordered in electron-density maps. Movements of other residues in the active site were shown to be related to the four displaced regions of the polypeptide chain. Their implied synergy suggests that DLH may be able to accommodate and catalyse a range of compounds unrelated to the natural substrate owing to an inherent coordinated flexibility in its overall structure. Implications for mechanism and further engineering studies are discussed.
 ==About this Structure==
-GGV is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida]. Active as [http://en.wikipedia.org/wiki/Carboxymethylenebutenolidase Carboxymethylenebutenolidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.1.45 3.1.1.45] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1GGV OCA].
+GGV is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudomonas_putida Pseudomonas putida]. Active as [http://en.wikipedia.org/wiki/Carboxymethylenebutenolidase Carboxymethylenebutenolidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.1.1.45 3.1.1.45] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1GGV OCA].
 ==Reference==
@@ Line 14: / Line 14: @@
 [[Category: Pseudomonas putida]]
 [[Category: Single protein]]
-[[Category: Barton, J.D.]]
+[[Category: Barton, J D.]]
-[[Category: Carr, P.D.]]
+[[Category: Carr, P D.]]
-[[Category: Edwards, K.J.]]
+[[Category: Edwards, K J.]]
-[[Category: Ewart, G.D.]]
+[[Category: Ewart, G D.]]
-[[Category: Ollis, D.L.]]
+[[Category: Ollis, D L.]]
 [[Category: Robinson, A.]]
 [[Category: alpha/beta hydrolase fold]]
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 [[Category: pseudomonas putida (pac27)]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 16:02:47 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:50:00 2008''