2e0w: Difference between revisions

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[[Image:2e0w.gif|left|200px]]<br /><applet load="2e0w" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:2e0w.gif|left|200px]]<br /><applet load="2e0w" size="350" color="white" frame="true" align="right" spinBox="true"

caption="2e0w, resolution 2.55Å" />

'''T391A precursor mutant protein of gamma-Glutamyltranspeptidase from Escherichia coli'''<br />

==Overview==

Gamma-glutamyltranspeptidase (GGT) is a heterodimic enzyme that is, generated from the precursor protein through posttranslational processing, and catalyzes the hydrolysis of gamma-glutamyl bonds in gamma-glutamyl, compounds such as glutathione and/or the transfer of the gamma-glutamyl, group to other amino acids and peptides. We have determined the crystal, structure of GGT from Escherichia coli K-12 at 1.95 A resolution. GGT has, a stacked alphabetabetaalpha fold comprising the large and small subunits, similar to the folds seen in members of the N-terminal nucleophile, hydrolase superfamily. The active site Thr-391, the N-terminal residue of, the small subunit, is located in the groove, from which the pocket for, gamma-glutamyl moiety binding follows. We have further determined the, structure of the gamma-glutamyl-enzyme intermediate trapped by flash, cooling the GGT crystal soaked in glutathione solution and the structure, of GGT in complex with l-glutamate. These structures revealed how the, gamma-glutamyl moiety and l-glutamate are recognized by the enzyme. A, water molecule was seen on the carbonyl carbon of the, gamma-glutamyl-Thr-391 Ogamma bond in the intermediate that is to be, hydrolyzed. Notably the residues essential for GGT activity (Arg-114, Asp-433, Ser-462, and Ser-463 in E. coli GGT) shown by site-directed, mutagenesis of human GGT are all involved in the binding of the, gamma-glutamyl moiety. The structure of E. coli GGT presented here, together with sequence alignment of GGTs, may be applicable to interpret, the biochemical and genetic data of other GGTs.

Gamma-glutamyltranspeptidase (GGT) is a heterodimic enzyme that is generated from the precursor protein through posttranslational processing and catalyzes the hydrolysis of gamma-glutamyl bonds in gamma-glutamyl compounds such as glutathione and/or the transfer of the gamma-glutamyl group to other amino acids and peptides. We have determined the crystal structure of GGT from Escherichia coli K-12 at 1.95 A resolution. GGT has a stacked alphabetabetaalpha fold comprising the large and small subunits, similar to the folds seen in members of the N-terminal nucleophile hydrolase superfamily. The active site Thr-391, the N-terminal residue of the small subunit, is located in the groove, from which the pocket for gamma-glutamyl moiety binding follows. We have further determined the structure of the gamma-glutamyl-enzyme intermediate trapped by flash cooling the GGT crystal soaked in glutathione solution and the structure of GGT in complex with l-glutamate. These structures revealed how the gamma-glutamyl moiety and l-glutamate are recognized by the enzyme. A water molecule was seen on the carbonyl carbon of the gamma-glutamyl-Thr-391 Ogamma bond in the intermediate that is to be hydrolyzed. Notably the residues essential for GGT activity (Arg-114, Asp-433, Ser-462, and Ser-463 in E. coli GGT) shown by site-directed mutagenesis of human GGT are all involved in the binding of the gamma-glutamyl moiety. The structure of E. coli GGT presented here, together with sequence alignment of GGTs, may be applicable to interpret the biochemical and genetic data of other GGTs.

==About this Structure==

2E0W is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli_k12 Escherichia coli k12]. Active as [http://en.wikipedia.org/wiki/Gamma-glutamyltransferase Gamma-glutamyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.3.2.2 2.3.2.2] Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=2E0W OCA].

2E0W is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli_k12 Escherichia coli k12]. Active as [http://en.wikipedia.org/wiki/Gamma-glutamyltransferase Gamma-glutamyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.3.2.2 2.3.2.2] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2E0W OCA].

==Reference==

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[[Category: precursor]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Wed Nov~~ 21 09:53:~~58 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:04:41 2008''

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@@ Line 1: / Line 1: @@
-[[Image:2e0w.gif|left|200px]]<br /><applet load="2e0w" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:2e0w.gif|left|200px]]<br /><applet load="2e0w" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="2e0w, resolution 2.55&Aring;" />
 '''T391A precursor mutant protein of gamma-Glutamyltranspeptidase from Escherichia coli'''<br />
 ==Overview==
-Gamma-glutamyltranspeptidase (GGT) is a heterodimic enzyme that is, generated from the precursor protein through posttranslational processing, and catalyzes the hydrolysis of gamma-glutamyl bonds in gamma-glutamyl, compounds such as glutathione and/or the transfer of the gamma-glutamyl, group to other amino acids and peptides. We have determined the crystal, structure of GGT from Escherichia coli K-12 at 1.95 A resolution. GGT has, a stacked alphabetabetaalpha fold comprising the large and small subunits, similar to the folds seen in members of the N-terminal nucleophile, hydrolase superfamily. The active site Thr-391, the N-terminal residue of, the small subunit, is located in the groove, from which the pocket for, gamma-glutamyl moiety binding follows. We have further determined the, structure of the gamma-glutamyl-enzyme intermediate trapped by flash, cooling the GGT crystal soaked in glutathione solution and the structure, of GGT in complex with l-glutamate. These structures revealed how the, gamma-glutamyl moiety and l-glutamate are recognized by the enzyme. A, water molecule was seen on the carbonyl carbon of the, gamma-glutamyl-Thr-391 Ogamma bond in the intermediate that is to be, hydrolyzed. Notably the residues essential for GGT activity (Arg-114, Asp-433, Ser-462, and Ser-463 in E. coli GGT) shown by site-directed, mutagenesis of human GGT are all involved in the binding of the, gamma-glutamyl moiety. The structure of E. coli GGT presented here, together with sequence alignment of GGTs, may be applicable to interpret, the biochemical and genetic data of other GGTs.
+Gamma-glutamyltranspeptidase (GGT) is a heterodimic enzyme that is generated from the precursor protein through posttranslational processing and catalyzes the hydrolysis of gamma-glutamyl bonds in gamma-glutamyl compounds such as glutathione and/or the transfer of the gamma-glutamyl group to other amino acids and peptides. We have determined the crystal structure of GGT from Escherichia coli K-12 at 1.95 A resolution. GGT has a stacked alphabetabetaalpha fold comprising the large and small subunits, similar to the folds seen in members of the N-terminal nucleophile hydrolase superfamily. The active site Thr-391, the N-terminal residue of the small subunit, is located in the groove, from which the pocket for gamma-glutamyl moiety binding follows. We have further determined the structure of the gamma-glutamyl-enzyme intermediate trapped by flash cooling the GGT crystal soaked in glutathione solution and the structure of GGT in complex with l-glutamate. These structures revealed how the gamma-glutamyl moiety and l-glutamate are recognized by the enzyme. A water molecule was seen on the carbonyl carbon of the gamma-glutamyl-Thr-391 Ogamma bond in the intermediate that is to be hydrolyzed. Notably the residues essential for GGT activity (Arg-114, Asp-433, Ser-462, and Ser-463 in E. coli GGT) shown by site-directed mutagenesis of human GGT are all involved in the binding of the gamma-glutamyl moiety. The structure of E. coli GGT presented here, together with sequence alignment of GGTs, may be applicable to interpret the biochemical and genetic data of other GGTs.
 ==About this Structure==
-E0W is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli_k12 Escherichia coli k12]. Active as [http://en.wikipedia.org/wiki/Gamma-glutamyltransferase Gamma-glutamyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.3.2.2 2.3.2.2] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=2E0W OCA].
+E0W is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Escherichia_coli_k12 Escherichia coli k12]. Active as [http://en.wikipedia.org/wiki/Gamma-glutamyltransferase Gamma-glutamyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.3.2.2 2.3.2.2] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2E0W OCA].
 ==Reference==
@@ Line 23: / Line 23: @@
 [[Category: precursor]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 09:53:58 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 17:04:41 2008''