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Visualizing Intracellular Distribution and Activity of Core2 β(1,6)N-Acetylglucosaminyltransferase-I in Living Cells

  • Maëlle Prorok-Hamon
  • Sylvie Mathieu
  • Assou El-Battari
Part of the Methods in Molecular Biology book series (MIMB, volume 347)

Abstract

The core2 β(1,6)-N-acetylglucosaminyltransferase-I (C2GnT-I) is expressed by leukocytes and is involved in the synthesis of core2 O-glycans that carry sialyl-Lewis x (sLex) oligosaccharides. The core2-based sLex oligosaccharides (C2-O-sLex) have been demonstrated to be physiological selectin ligands that confer high affinity binding. The E-, P-, and L-selectins are adhesion proteins that direct leukocytes in the blood to lymphoid organs and sites of inflammation. They are also thought to be involved in the hematogenous dissemination of carcinoma cells expressing sialyl-Lewis glycans. Therefore, accumulation of data on structure-function relationships of this particular enzyme may represent an important part of investigations into pathologies involving selectins, such us inflammatory disorders and cancer progression. In this regard, studies of the intracellular distribution of C2GnT-I and its interaction with cognate substrates in vivo, as well as the knowledge of posttranslational modification (i.e., glycosylation, oligomerization, and proteolytic processing), may greatly aid in designing potential enzyme inhibitors. C2GnT-I fused to the green fluorescent protein is expressed to allow examination of the protein in living cells and to ease studies on structure-function relationships in vivo and in vitro.

Keywords

Sodium Dodecyl Sulfate Enhanced Green Fluorescent Protein Chinese Hamster Ovary Cell Enhanced Green Fluorescent Protein Gene Enhanced Green Fluorescent Protein Fluorescence 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Maëlle Prorok-Hamon
    • 1
  • Sylvie Mathieu
    • 1
  • Assou El-Battari
    • 1
    • 2
  1. 1.Facultée de MédecineINSERM U-559MarseilleFrance
  2. 2.Université de ProvenceMarseilleFrance

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