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Crystal Structure of Rabbit Muscle Glycogenin

OverviewOverview

Glycogen is an important storage reserve of glucose present in many organisms, from bacteria to humans. Its biosynthesis is initiated by a specialized protein, glycogenin, which has the unusual property of transferring glucose from UDP-glucose to form an oligosaccharide covalently attached to itself at Tyr194. Glycogen synthase and the branching enzyme complete the synthesis of the polysaccharide. The structure of glycogenin was solved in two different crystal forms. Tetragonal crystals contained a pentamer of dimers in the asymmetric unit arranged in an improper non-crystallographic 10-fold relationship, and orthorhombic crystals contained a monomer in the asymmetric unit that is arranged about a 2-fold crystallographic axis to form a dimer. The structure was first solved to 3.4 A using the tetragonal crystal form and a three-wavelength Se-Met multi-wavelength anomalous diffraction (MAD) experiment. Subsequently, an apo-enzyme structure and a complex between glycogenin and UDP-glucose/Mn2+ were solved by molecular replacement to 1.9 A using the orthorhombic crystal form. Glycogenin contains a conserved DxD motif and an N-terminal beta-alpha-beta Rossmann-like fold that are common to the nucleotide-binding domains of most glycosyltransferases. Although sequence identity amongst glycosyltransferases is minimal, the overall folds are similar. In all of these enzymes, the DxD motif is essential for coordination of the catalytic divalent cation, most commonly Mn2+. We propose a mechanism in which the Mn2+ that associates with the UDP-glucose molecule functions as a Lewis acid to stabilize the leaving group UDP and to facilitate the transfer of the glucose moiety to an intermediate nucleophilic acceptor in the enzyme active site, most likely Asp162. Following transient transfer to Asp162, the glucose moiety is then delivered to the final acceptor, either directly to Tyr194 or to glucose residues already attached to Tyr194. The positioning of the bound UDP-glucose far from Tyr194 in the glycogenin structure raises questions as to the mechanism for the attachment of the first glucose residues. Possibly the initial glucosylation is via inter-dimeric catalysis with an intra-molecular mechanism employed later in oligosaccharide synthesis.

About this StructureAbout this Structure

1LL0 is a Single protein structure of sequence from Oryctolagus cuniculus. Active as Glycogenin glucosyltransferase, with EC number 2.4.1.186 Full crystallographic information is available from OCA.

ReferenceReference

Crystal structure of the autocatalytic initiator of glycogen biosynthesis, glycogenin., Gibbons BJ, Roach PJ, Hurley TD, J Mol Biol. 2002 May 31;319(2):463-77. PMID:12051921

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-[[Image:1ll0.gif|left|200px]]<br /><applet load="1ll0" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1ll0.gif|left|200px]]<br /><applet load="1ll0" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1ll0, resolution 3.43&Aring;" />
 '''Crystal Structure of Rabbit Muscle Glycogenin'''<br />
 ==Overview==
-Glycogen is an important storage reserve of glucose present in many, organisms, from bacteria to humans. Its biosynthesis is initiated by a, specialized protein, glycogenin, which has the unusual property of, transferring glucose from UDP-glucose to form an oligosaccharide, covalently attached to itself at Tyr194. Glycogen synthase and the, branching enzyme complete the synthesis of the polysaccharide. The, structure of glycogenin was solved in two different crystal forms., Tetragonal crystals contained a pentamer of dimers in the asymmetric unit, arranged in an improper non-crystallographic 10-fold relationship, and, orthorhombic crystals contained a monomer in the asymmetric unit that is, arranged about a 2-fold crystallographic axis to form a dimer. The, structure was first solved to 3.4 A using the tetragonal crystal form and, a three-wavelength Se-Met multi-wavelength anomalous diffraction (MAD), experiment. Subsequently, an apo-enzyme structure and a complex between, glycogenin and UDP-glucose/Mn2+ were solved by molecular replacement to, 1.9 A using the orthorhombic crystal form. Glycogenin contains a conserved, DxD motif and an N-terminal beta-alpha-beta Rossmann-like fold that are, common to the nucleotide-binding domains of most glycosyltransferases., Although sequence identity amongst glycosyltransferases is minimal, the, overall folds are similar. In all of these enzymes, the DxD motif is, essential for coordination of the catalytic divalent cation, most commonly, Mn2+. We propose a mechanism in which the Mn2+ that associates with the, UDP-glucose molecule functions as a Lewis acid to stabilize the leaving, group UDP and to facilitate the transfer of the glucose moiety to an, intermediate nucleophilic acceptor in the enzyme active site, most likely, Asp162. Following transient transfer to Asp162, the glucose moiety is then, delivered to the final acceptor, either directly to Tyr194 or to glucose, residues already attached to Tyr194. The positioning of the bound, UDP-glucose far from Tyr194 in the glycogenin structure raises questions, as to the mechanism for the attachment of the first glucose residues., Possibly the initial glucosylation is via inter-dimeric catalysis with an, intra-molecular mechanism employed later in oligosaccharide synthesis.
+Glycogen is an important storage reserve of glucose present in many organisms, from bacteria to humans. Its biosynthesis is initiated by a specialized protein, glycogenin, which has the unusual property of transferring glucose from UDP-glucose to form an oligosaccharide covalently attached to itself at Tyr194. Glycogen synthase and the branching enzyme complete the synthesis of the polysaccharide. The structure of glycogenin was solved in two different crystal forms. Tetragonal crystals contained a pentamer of dimers in the asymmetric unit arranged in an improper non-crystallographic 10-fold relationship, and orthorhombic crystals contained a monomer in the asymmetric unit that is arranged about a 2-fold crystallographic axis to form a dimer. The structure was first solved to 3.4 A using the tetragonal crystal form and a three-wavelength Se-Met multi-wavelength anomalous diffraction (MAD) experiment. Subsequently, an apo-enzyme structure and a complex between glycogenin and UDP-glucose/Mn2+ were solved by molecular replacement to 1.9 A using the orthorhombic crystal form. Glycogenin contains a conserved DxD motif and an N-terminal beta-alpha-beta Rossmann-like fold that are common to the nucleotide-binding domains of most glycosyltransferases. Although sequence identity amongst glycosyltransferases is minimal, the overall folds are similar. In all of these enzymes, the DxD motif is essential for coordination of the catalytic divalent cation, most commonly Mn2+. We propose a mechanism in which the Mn2+ that associates with the UDP-glucose molecule functions as a Lewis acid to stabilize the leaving group UDP and to facilitate the transfer of the glucose moiety to an intermediate nucleophilic acceptor in the enzyme active site, most likely Asp162. Following transient transfer to Asp162, the glucose moiety is then delivered to the final acceptor, either directly to Tyr194 or to glucose residues already attached to Tyr194. The positioning of the bound UDP-glucose far from Tyr194 in the glycogenin structure raises questions as to the mechanism for the attachment of the first glucose residues. Possibly the initial glucosylation is via inter-dimeric catalysis with an intra-molecular mechanism employed later in oligosaccharide synthesis.
 ==About this Structure==
-LL0 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Oryctolagus_cuniculus Oryctolagus cuniculus]. Active as [http://en.wikipedia.org/wiki/Glycogenin_glucosyltransferase Glycogenin glucosyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.186 2.4.1.186] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1LL0 OCA].
+LL0 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Oryctolagus_cuniculus Oryctolagus cuniculus]. Active as [http://en.wikipedia.org/wiki/Glycogenin_glucosyltransferase Glycogenin glucosyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.186 2.4.1.186] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1LL0 OCA].
 ==Reference==
@@ Line 14: / Line 14: @@
 [[Category: Oryctolagus cuniculus]]
 [[Category: Single protein]]
-[[Category: Gibbons, B.J.]]
+[[Category: Gibbons, B J.]]
-[[Category: Hurley, T.D.]]
+[[Category: Hurley, T D.]]
-[[Category: Roach, P.J.]]
+[[Category: Roach, P J.]]
 [[Category: beta-alpha-beta rossman-like nucleotide binding fold]]
 [[Category: dxd motif]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Nov 20 20:38:30 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:45:56 2008''