X-ray Crystal Structure of Leukocyte Type Core 2 β1 ,6-N-Acetylglucosaminyltransferase.

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Title: X-ray Crystal Structure of Leukocyte Type Core 2 β1 ,6-N-Acetylglucosaminyltransferase.
Authors: Pak, John E.1, Arnoux, Pascal1, Sihong Zhou1, Sivarajah, Prashanth1, Satkunarajah, Malathy1, Xuekun Xing1, Rini, James M.1 james.rini@utoronto.ca
Source: Journal of Biological Chemistry. 9/8/2006, Vol. 281 Issue 36, p26693-26701. 9p. 6 Diagrams, 1 Chart.
Subjects: X-ray crystallography, Biosynthesis, Leukocytes, Glycosyltransferases, Glucans, Nucleosides, Biochemistry
Abstract: Leukocyte type core 2 β1,6-N-acetylglucosaminyltransferase (C2GnT-L) is a key enzyme in the biosynthesis of branched O-glycans. It is an inverting, metal ion-independent family 14 glycosyltransferase that catalyzes the formation of the core 2 O-glycan (Galβ1-3[GlcNAcβ1-6]GalNAc-O-Ser/Thr) from its donor and acceptor substrates, UDP-GIcNAc and the core 1 O-glycan (Galβ1-3GalNAc-O-Ser/Thr), respectively. Reported here are the x-ray crystal structures of murine C2GnT-L in the absence and presence of the acceptor substrate Galβ1-3GalNAc at 2.0 and 2.7 Å resolution, respectively. C2GnT-L was found to possess the GT-A fold; however, it lacks the characteristic metal ion binding DXD motif. The Galβ1-3GalNAc complex defines the determinants of acceptor substrate binding and shows that Glu-320 corresponds to the structurally conserved catalytic base found in other inverting GT-A fold glycosyltransferases. Comparison of the C2GnT-L structure with that of other GT-A fold glycosyltransferases further suggests that Arg-378 and Lys-401 serve to electrostatically stabilize the nucleoside disphosphate leaving group, a role normally played by metal ion in GT-A structures. The use of basic amino acid side chains in this way is strikingly similar to that seen in a number of metal ion-independent GT-B fold glycosyltransferases and suggests a convergence of catalytic mechanism shared by both GT-A and GT-B fold glycosyltransferases. [ABSTRACT FROM AUTHOR]
Copyright of Journal of Biological Chemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: X-ray Crystal Structure of Leukocyte Type Core 2 β1 ,6-N-Acetylglucosaminyltransferase.
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  Data: <searchLink fieldCode="AR" term="%22Pak%2C+John+E%2E%22">Pak, John E.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Arnoux%2C+Pascal%22">Arnoux, Pascal</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Sihong+Zhou%22">Sihong Zhou</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Sivarajah%2C+Prashanth%22">Sivarajah, Prashanth</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Satkunarajah%2C+Malathy%22">Satkunarajah, Malathy</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Xuekun+Xing%22">Xuekun Xing</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Rini%2C+James+M%2E%22">Rini, James M.</searchLink><relatesTo>1</relatesTo><i> james.rini@utoronto.ca</i>
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  Data: <searchLink fieldCode="JN" term="%22Journal+of+Biological+Chemistry%22">Journal of Biological Chemistry</searchLink>. 9/8/2006, Vol. 281 Issue 36, p26693-26701. 9p. 6 Diagrams, 1 Chart.
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  Data: <searchLink fieldCode="DE" term="%22X-ray+crystallography%22">X-ray crystallography</searchLink><br /><searchLink fieldCode="DE" term="%22Biosynthesis%22">Biosynthesis</searchLink><br /><searchLink fieldCode="DE" term="%22Leukocytes%22">Leukocytes</searchLink><br /><searchLink fieldCode="DE" term="%22Glycosyltransferases%22">Glycosyltransferases</searchLink><br /><searchLink fieldCode="DE" term="%22Glucans%22">Glucans</searchLink><br /><searchLink fieldCode="DE" term="%22Nucleosides%22">Nucleosides</searchLink><br /><searchLink fieldCode="DE" term="%22Biochemistry%22">Biochemistry</searchLink>
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  Data: Leukocyte type core 2 β1,6-N-acetylglucosaminyltransferase (C2GnT-L) is a key enzyme in the biosynthesis of branched O-glycans. It is an inverting, metal ion-independent family 14 glycosyltransferase that catalyzes the formation of the core 2 O-glycan (Galβ1-3[GlcNAcβ1-6]GalNAc-O-Ser/Thr) from its donor and acceptor substrates, UDP-GIcNAc and the core 1 O-glycan (Galβ1-3GalNAc-O-Ser/Thr), respectively. Reported here are the x-ray crystal structures of murine C2GnT-L in the absence and presence of the acceptor substrate Galβ1-3GalNAc at 2.0 and 2.7 Å resolution, respectively. C2GnT-L was found to possess the GT-A fold; however, it lacks the characteristic metal ion binding DXD motif. The Galβ1-3GalNAc complex defines the determinants of acceptor substrate binding and shows that Glu-320 corresponds to the structurally conserved catalytic base found in other inverting GT-A fold glycosyltransferases. Comparison of the C2GnT-L structure with that of other GT-A fold glycosyltransferases further suggests that Arg-378 and Lys-401 serve to electrostatically stabilize the nucleoside disphosphate leaving group, a role normally played by metal ion in GT-A structures. The use of basic amino acid side chains in this way is strikingly similar to that seen in a number of metal ion-independent GT-B fold glycosyltransferases and suggests a convergence of catalytic mechanism shared by both GT-A and GT-B fold glycosyltransferases. [ABSTRACT FROM AUTHOR]
– Name: AbstractSuppliedCopyright
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  Data: <i>Copyright of Journal of Biological Chemistry is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1074/jbc.M603534200
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      – Code: eng
        Text: English
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        PageCount: 9
        StartPage: 26693
    Subjects:
      – SubjectFull: X-ray crystallography
        Type: general
      – SubjectFull: Biosynthesis
        Type: general
      – SubjectFull: Leukocytes
        Type: general
      – SubjectFull: Glycosyltransferases
        Type: general
      – SubjectFull: Glucans
        Type: general
      – SubjectFull: Nucleosides
        Type: general
      – SubjectFull: Biochemistry
        Type: general
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      – TitleFull: X-ray Crystal Structure of Leukocyte Type Core 2 β1 ,6-N-Acetylglucosaminyltransferase.
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            NameFull: Arnoux, Pascal
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            NameFull: Sihong Zhou
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            NameFull: Sivarajah, Prashanth
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            NameFull: Satkunarajah, Malathy
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              Text: 9/8/2006
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              Y: 2006
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