1v83

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(New page: 200px<br /> <applet load="1v83" size="450" color="white" frame="true" align="right" spinBox="true" caption="1v83, resolution 1.9&Aring;" /> '''Crystal structure of...)
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'''Crystal structure of human GlcAT-P in complex with Udp and Mn2+'''<br />
'''Crystal structure of human GlcAT-P in complex with Udp and Mn2+'''<br />
==Overview==
==Overview==
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The HNK-1 carbohydrate epitope is found on many neural cell adhesion, molecules. Its structure is characterized by a terminal sulfated, glucuronyl acid. The glucuronyltransferases, GlcAT-P and GlcAT-S, are, involved in the biosynthesis of the HNK-1 epitope, GlcAT-P as the major, enzyme. We overexpressed and purified the recombinant human GlcAT-P from, Escherichia coli. Analysis of its enzymatic activity showed that it, catalyzed the transfer reaction for N-acetyllactosamine (Galbeta1-4GlcNAc), but not lacto-N-biose (Galbeta1-3GlcNAc) as an acceptor substrate., Subsequently, we determined the first x-ray crystal structures of human, GlcAT-P, in the absence and presence of a donor substrate product UDP, catalytic Mn(2+), and an acceptor substrate analogue N-acetyllactosamine, (Galbeta1-4GlcNAc) or an asparagine-linked biantennary nonasaccharide. The, asymmetric unit contains two independent molecules. Each molecule is an, alpha/beta protein with two regions that constitute the donor and acceptor, substrate binding sites. The UDP moiety of donor nucleotide sugar is, recognized by conserved amino acid residues including a DXD motif, (Asp(195)-Asp(196)-Asp(197)). Other conserved amino acid residues interact, with the terminal galactose moiety of the acceptor substrate. In addition, Val(320) and Asn(321), which are located on the C-terminal long loop from, a neighboring molecule, and Phe(245) contribute to the interaction with, GlcNAc moiety. These three residues play a key role in establishing the, acceptor substrate specificity.
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The HNK-1 carbohydrate epitope is found on many neural cell adhesion molecules. Its structure is characterized by a terminal sulfated glucuronyl acid. The glucuronyltransferases, GlcAT-P and GlcAT-S, are involved in the biosynthesis of the HNK-1 epitope, GlcAT-P as the major enzyme. We overexpressed and purified the recombinant human GlcAT-P from Escherichia coli. Analysis of its enzymatic activity showed that it catalyzed the transfer reaction for N-acetyllactosamine (Galbeta1-4GlcNAc) but not lacto-N-biose (Galbeta1-3GlcNAc) as an acceptor substrate. Subsequently, we determined the first x-ray crystal structures of human GlcAT-P, in the absence and presence of a donor substrate product UDP, catalytic Mn(2+), and an acceptor substrate analogue N-acetyllactosamine (Galbeta1-4GlcNAc) or an asparagine-linked biantennary nonasaccharide. The asymmetric unit contains two independent molecules. Each molecule is an alpha/beta protein with two regions that constitute the donor and acceptor substrate binding sites. The UDP moiety of donor nucleotide sugar is recognized by conserved amino acid residues including a DXD motif (Asp(195)-Asp(196)-Asp(197)). Other conserved amino acid residues interact with the terminal galactose moiety of the acceptor substrate. In addition, Val(320) and Asn(321), which are located on the C-terminal long loop from a neighboring molecule, and Phe(245) contribute to the interaction with GlcNAc moiety. These three residues play a key role in establishing the acceptor substrate specificity.
==About this Structure==
==About this Structure==
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1V83 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with MN, TLA and UDP as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Galactosylgalactosylxylosylprotein_3-beta-glucuronosyltransferase Galactosylgalactosylxylosylprotein 3-beta-glucuronosyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.135 2.4.1.135] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1V83 OCA].
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1V83 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Homo_sapiens Homo sapiens] with <scene name='pdbligand=MN:'>MN</scene>, <scene name='pdbligand=TLA:'>TLA</scene> and <scene name='pdbligand=UDP:'>UDP</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Galactosylgalactosylxylosylprotein_3-beta-glucuronosyltransferase Galactosylgalactosylxylosylprotein 3-beta-glucuronosyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.135 2.4.1.135] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1V83 OCA].
==Reference==
==Reference==
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[[Category: transferase]]
[[Category: transferase]]
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''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Mon Nov 12 19:42:07 2007''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:32:28 2008''

Revision as of 13:32, 21 February 2008

Image:1v83.gif

1v83, resolution 1.9Å

Drag the structure with the mouse to rotate

Crystal structure of human GlcAT-P in complex with Udp and Mn2+

Overview

The HNK-1 carbohydrate epitope is found on many neural cell adhesion molecules. Its structure is characterized by a terminal sulfated glucuronyl acid. The glucuronyltransferases, GlcAT-P and GlcAT-S, are involved in the biosynthesis of the HNK-1 epitope, GlcAT-P as the major enzyme. We overexpressed and purified the recombinant human GlcAT-P from Escherichia coli. Analysis of its enzymatic activity showed that it catalyzed the transfer reaction for N-acetyllactosamine (Galbeta1-4GlcNAc) but not lacto-N-biose (Galbeta1-3GlcNAc) as an acceptor substrate. Subsequently, we determined the first x-ray crystal structures of human GlcAT-P, in the absence and presence of a donor substrate product UDP, catalytic Mn(2+), and an acceptor substrate analogue N-acetyllactosamine (Galbeta1-4GlcNAc) or an asparagine-linked biantennary nonasaccharide. The asymmetric unit contains two independent molecules. Each molecule is an alpha/beta protein with two regions that constitute the donor and acceptor substrate binding sites. The UDP moiety of donor nucleotide sugar is recognized by conserved amino acid residues including a DXD motif (Asp(195)-Asp(196)-Asp(197)). Other conserved amino acid residues interact with the terminal galactose moiety of the acceptor substrate. In addition, Val(320) and Asn(321), which are located on the C-terminal long loop from a neighboring molecule, and Phe(245) contribute to the interaction with GlcNAc moiety. These three residues play a key role in establishing the acceptor substrate specificity.

About this Structure

1V83 is a Single protein structure of sequence from Homo sapiens with , and as ligands. Active as Galactosylgalactosylxylosylprotein 3-beta-glucuronosyltransferase, with EC number 2.4.1.135 Full crystallographic information is available from OCA.

Reference

Structural basis for acceptor substrate recognition of a human glucuronyltransferase, GlcAT-P, an enzyme critical in the biosynthesis of the carbohydrate epitope HNK-1., Kakuda S, Shiba T, Ishiguro M, Tagawa H, Oka S, Kajihara Y, Kawasaki T, Wakatsuki S, Kato R, J Biol Chem. 2004 May 21;279(21):22693-703. Epub 2004 Mar 1. PMID:14993226

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