1oxw: Difference between revisions

Line 1:

[[Image:1oxw.gif|left|200px]]<br /><applet load="1oxw" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:1oxw.gif|left|200px]]<br /><applet load="1oxw" size="350" color="white" frame="true" align="right" spinBox="true"

caption="1oxw, resolution 2.20Å" />

'''The Crystal Structure of SeMet Patatin'''<br />

==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.

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 [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://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=1OXW OCA].

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==

Line 13:

[[Category: Single protein]]

[[Category: Solanum cardiophyllum]]

[[Category: Alibhai, M.F.]]

[[Category: Alibhai, M F.]]

[[Category: Brown, S.M.]]

[[Category: Brown, S M.]]

[[Category: Krieger, E.]]

[[Category: Moshiri, F.]]

[[Category: Pershing, J.C.]]

[[Category: Pershing, J C.]]

[[Category: Purcell, J.P.]]

[[Category: Purcell, J P.]]

[[Category: Rydel, T.J.]]

[[Category: Rydel, T J.]]

[[Category: Stallings, W.C.]]

[[Category: Stallings, W C.]]

[[Category: Williams, J.M.]]

[[Category: Williams, J M.]]

[[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]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Tue Nov 20 23~~:17:~~02 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:22:54 2008''

Navigation menu

Page actions

Page actions

Personal tools

Navigation

Search

Tools

@@ Line 1: / Line 1: @@
-[[Image:1oxw.gif|left|200px]]<br /><applet load="1oxw" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1oxw.gif|left|200px]]<br /><applet load="1oxw" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1oxw, resolution 2.20&Aring;" />
 '''The Crystal Structure of SeMet Patatin'''<br />
 ==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.
+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==
-OXW 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://ispc.weizmann.ac.il/oca-bin/ocashort?id=1OXW OCA].
+OXW 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==
@@ Line 13: / Line 13: @@
 [[Category: Single protein]]
 [[Category: Solanum cardiophyllum]]
-[[Category: Alibhai, M.F.]]
+[[Category: Alibhai, M F.]]
-[[Category: Brown, S.M.]]
+[[Category: Brown, S M.]]
 [[Category: Krieger, E.]]
 [[Category: Moshiri, F.]]
-[[Category: Pershing, J.C.]]
+[[Category: Pershing, J C.]]
-[[Category: Purcell, J.P.]]
+[[Category: Purcell, J P.]]
-[[Category: Rydel, T.J.]]
+[[Category: Rydel, T J.]]
-[[Category: Stallings, W.C.]]
+[[Category: Stallings, W C.]]
-[[Category: Williams, J.M.]]
+[[Category: Williams, J M.]]
 [[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]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 23:17:02 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:22:54 2008''