2plq: Difference between revisions

Revision as of 19:30, 21 February 2008

Crystal structure of the amidase from geobacillus pallidus RAPc8

OverviewOverview

The amidase from Geobacillus pallidus RAPc8, a moderate thermophile, is a member of the nitrilase enzyme superfamily. It converts amides to the corresponding acids and ammonia and has application as an industrial catalyst. RAPc8 amidase has been cloned and functionally expressed in Escherichia coli and has been purified by heat treatment and a number of chromatographic steps. The enzyme was crystallized using the hanging-drop vapour-diffusion method. Crystals produced in the presence of 1.2 M sodium citrate, 400 mM NaCl, 100 mM sodium acetate pH 5.6 were selected for X-ray diffraction studies. A data set having acceptable statistics to 1.96 A resolution was collected under cryoconditions using an in-house X-ray source. The space group was determined to be primitive cubic P4(2)32, with unit-cell parameter a = 130.49 (+/-0.05) A. The structure was solved by molecular replacement using the backbone of the hypothetical protein PH0642 from Pyrococcus horikoshii (PDB code 1j31) with all non-identical side chains substituted with alanine as a probe. There is one subunit per asymmetric unit. The subunits are packed as trimers of dimers with D3 point-group symmetry around the threefold axis in such a way that the dimer interface seen in the homologues is preserved.

About this StructureAbout this Structure

2PLQ is a Single protein structure of sequence from Geobacillus pallidus. Active as Amidase, with EC number 3.5.1.4 Full crystallographic information is available from OCA.

ReferenceReference

The quaternary structure of the amidase from Geobacillus pallidus RAPc8 is revealed by its crystal packing., Agarkar VB, Kimani SW, Cowan DA, Sayed MF, Sewell BT, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Dec 1;62(Pt, 12):1174-8. Epub 2006 Nov 4. PMID:17142891

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 ==Overview==
-The amidase from Geobacillus pallidus RAPc8, a moderate thermophile, is a, member of the nitrilase enzyme superfamily. It converts amides to the, corresponding acids and ammonia and has application as an industrial, catalyst. RAPc8 amidase has been cloned and functionally expressed in, Escherichia coli and has been purified by heat treatment and a number of, chromatographic steps. The enzyme was crystallized using the hanging-drop, vapour-diffusion method. Crystals produced in the presence of 1.2 M sodium, citrate, 400 mM NaCl, 100 mM sodium acetate pH 5.6 were selected for X-ray, diffraction studies. A data set having acceptable statistics to 1.96 A, resolution was collected under cryoconditions using an in-house X-ray, source. The space group was determined to be primitive cubic P4(2)32, with, unit-cell parameter a = 130.49 (+/-0.05) A. The structure was solved by, molecular replacement using the backbone of the hypothetical protein, PH0642 from Pyrococcus horikoshii (PDB code 1j31) with all non-identical, side chains substituted with alanine as a probe. There is one subunit per, asymmetric unit. The subunits are packed as trimers of dimers with D3, point-group symmetry around the threefold axis in such a way that the, dimer interface seen in the homologues is preserved.
+The amidase from Geobacillus pallidus RAPc8, a moderate thermophile, is a member of the nitrilase enzyme superfamily. It converts amides to the corresponding acids and ammonia and has application as an industrial catalyst. RAPc8 amidase has been cloned and functionally expressed in Escherichia coli and has been purified by heat treatment and a number of chromatographic steps. The enzyme was crystallized using the hanging-drop vapour-diffusion method. Crystals produced in the presence of 1.2 M sodium citrate, 400 mM NaCl, 100 mM sodium acetate pH 5.6 were selected for X-ray diffraction studies. A data set having acceptable statistics to 1.96 A resolution was collected under cryoconditions using an in-house X-ray source. The space group was determined to be primitive cubic P4(2)32, with unit-cell parameter a = 130.49 (+/-0.05) A. The structure was solved by molecular replacement using the backbone of the hypothetical protein PH0642 from Pyrococcus horikoshii (PDB code 1j31) with all non-identical side chains substituted with alanine as a probe. There is one subunit per asymmetric unit. The subunits are packed as trimers of dimers with D3 point-group symmetry around the threefold axis in such a way that the dimer interface seen in the homologues is preserved.
 ==About this Structure==
@@ Line 10: / Line 10: @@
 ==Reference==
-The quaternary structure of the amidase from Geobacillus pallidus RAPc8 is revealed by its crystal packing., Agarkar VB, Kimani SW, Cowan DA, Sayed MF, Sewell BT, Acta Crystallograph Sect F Struct Biol Cryst Commun. 2006 Dec 1;62(Pt, 12):1174-8. Epub 2006 Nov 4. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17142891 17142891]
+The quaternary structure of the amidase from Geobacillus pallidus RAPc8 is revealed by its crystal packing., Agarkar VB, Kimani SW, Cowan DA, Sayed MF, Sewell BT, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Dec 1;62(Pt, 12):1174-8. Epub 2006 Nov 4. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17142891 17142891]
 [[Category: Amidase]]
 [[Category: Geobacillus pallidus]]
 [[Category: Single protein]]
-[[Category: Agarkar, V.B.]]
+[[Category: Agarkar, V B.]]
-[[Category: Cowan, D.A.]]
+[[Category: Cowan, D A.]]
-[[Category: Kimani, S.W.]]
+[[Category: Kimani, S W.]]
-[[Category: Sayed, M.F.]]
+[[Category: Sayed, M F.]]
-[[Category: Sewell, B.T.]]
+[[Category: Sewell, B T.]]
 [[Category: alpha-beta-beta-alpha]]
 [[Category: nitrilase fold]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 15:29:28 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:30:47 2008''