1oxw
From Proteopedia
(New page: 200px<br /><applet load="1oxw" size="450" color="white" frame="true" align="right" spinBox="true" caption="1oxw, resolution 2.20Å" /> '''The Crystal Structur...) |
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| - | [[Image:1oxw.gif|left|200px]]<br /><applet load="1oxw" size=" | + | [[Image:1oxw.gif|left|200px]]<br /><applet load="1oxw" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1oxw, resolution 2.20Å" /> | caption="1oxw, resolution 2.20Å" /> | ||
'''The Crystal Structure of SeMet Patatin'''<br /> | '''The Crystal Structure of SeMet Patatin'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Patatin is a nonspecific lipid acyl hydrolase that accounts for | + | Patatin is a nonspecific lipid acyl hydrolase that accounts for approximately 40% of the total soluble protein in mature potato tubers, and it has potent insecticidal activity against the corn rootworm. We determined the X-ray crystal structure of a His-tagged variant of an isozyme of patatin, Pat17, to 2.2 A resolution, employing SeMet multiwavelength anomalous dispersion (MAD) phasing methods. The patatin crystal structure has three molecules in the asymmetric unit, an R-factor of 22.0%, and an R(free) of 27.2% (for 10% of the data not included in the refinement) and includes 498 water molecules. The structure notably revealed that patatin has a Ser-Asp catalytic dyad and an active site like that of human cytosolic phospholipase A(2) (cPLA(2)) [Dessen, A., et al. (1999) Cell 97, 349-360]. In addition, patatin has a folding topology related to that of the catalytic domain of cPLA(2) and unlike the canonical alpha/beta-hydrolase fold. The structure confirms our site-directed mutagenesis and bioactivity data that initially suggested patatin possessed a Ser-Asp catalytic dyad. Alanine-scanning mutagenesis revealed that Ser77 and Asp215 were critical for both esterase and bioactivity, consistent with prior work implicating a Ser residue [Strickland, J. H., et al. (1995) Plant Physiol. 109, 667-674] and a Ser-Asp dyad [Hirschberg, H. J. H. B., et al. (2001) Eur. J. Biochem. 268, 5037-5044] in patatin's catalytic activity. The crystal structure aids the understanding of other structure-function relationships in patatin. Patatin does not display interfacial activation, a hallmark feature of lipases, and this is likely due to the fact that it lacks a flexible lid that can shield the active site. |
==About this Structure== | ==About this Structure== | ||
| - | 1OXW is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Solanum_cardiophyllum Solanum cardiophyllum]. Full crystallographic information is available from [http:// | + | 1OXW is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Solanum_cardiophyllum Solanum cardiophyllum]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1OXW OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Single protein]] | [[Category: Single protein]] | ||
[[Category: Solanum cardiophyllum]] | [[Category: Solanum cardiophyllum]] | ||
| - | [[Category: Alibhai, M | + | [[Category: Alibhai, M F.]] |
| - | [[Category: Brown, S | + | [[Category: Brown, S M.]] |
[[Category: Krieger, E.]] | [[Category: Krieger, E.]] | ||
[[Category: Moshiri, F.]] | [[Category: Moshiri, F.]] | ||
| - | [[Category: Pershing, J | + | [[Category: Pershing, J C.]] |
| - | [[Category: Purcell, J | + | [[Category: Purcell, J P.]] |
| - | [[Category: Rydel, T | + | [[Category: Rydel, T J.]] |
| - | [[Category: Stallings, W | + | [[Category: Stallings, W C.]] |
| - | [[Category: Williams, J | + | [[Category: Williams, J M.]] |
[[Category: alpha/beta class fold with approximately three layers]] | [[Category: alpha/beta class fold with approximately three layers]] | ||
[[Category: alpha/beta/alpha in content. possesses a central six-stranded beta sheet with alpha-helices front & back]] | [[Category: alpha/beta/alpha in content. possesses a central six-stranded beta sheet with alpha-helices front & back]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:22:54 2008'' |
Revision as of 12:22, 21 February 2008
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The Crystal Structure of SeMet Patatin
Overview
Patatin is a nonspecific lipid acyl hydrolase that accounts for approximately 40% of the total soluble protein in mature potato tubers, and it has potent insecticidal activity against the corn rootworm. We determined the X-ray crystal structure of a His-tagged variant of an isozyme of patatin, Pat17, to 2.2 A resolution, employing SeMet multiwavelength anomalous dispersion (MAD) phasing methods. The patatin crystal structure has three molecules in the asymmetric unit, an R-factor of 22.0%, and an R(free) of 27.2% (for 10% of the data not included in the refinement) and includes 498 water molecules. The structure notably revealed that patatin has a Ser-Asp catalytic dyad and an active site like that of human cytosolic phospholipase A(2) (cPLA(2)) [Dessen, A., et al. (1999) Cell 97, 349-360]. In addition, patatin has a folding topology related to that of the catalytic domain of cPLA(2) and unlike the canonical alpha/beta-hydrolase fold. The structure confirms our site-directed mutagenesis and bioactivity data that initially suggested patatin possessed a Ser-Asp catalytic dyad. Alanine-scanning mutagenesis revealed that Ser77 and Asp215 were critical for both esterase and bioactivity, consistent with prior work implicating a Ser residue [Strickland, J. H., et al. (1995) Plant Physiol. 109, 667-674] and a Ser-Asp dyad [Hirschberg, H. J. H. B., et al. (2001) Eur. J. Biochem. 268, 5037-5044] in patatin's catalytic activity. The crystal structure aids the understanding of other structure-function relationships in patatin. Patatin does not display interfacial activation, a hallmark feature of lipases, and this is likely due to the fact that it lacks a flexible lid that can shield the active site.
About this Structure
1OXW is a Single protein structure of sequence from Solanum cardiophyllum. Full crystallographic information is available from OCA.
Reference
The crystal structure, mutagenesis, and activity studies reveal that patatin is a lipid acyl hydrolase with a Ser-Asp catalytic dyad., Rydel TJ, Williams JM, Krieger E, Moshiri F, Stallings WC, Brown SM, Pershing JC, Purcell JP, Alibhai MF, Biochemistry. 2003 Jun 10;42(22):6696-708. PMID:12779324
Page seeded by OCA on Thu Feb 21 14:22:54 2008
Categories: Single protein | Solanum cardiophyllum | Alibhai, M F. | Brown, S M. | Krieger, E. | Moshiri, F. | Pershing, J C. | Purcell, J P. | Rydel, T J. | Stallings, W C. | Williams, J M. | Alpha/beta class fold with approximately three layers | Alpha/beta/alpha in content. possesses a central six-stranded beta sheet with alpha-helices front & back
