A cell-free assay for glycosylphosphatidylinositol anchoring in African trypanosomes. Demonstration of a transamidation reaction mechanism.

@article{Sharma1999ACA,
  title={A cell-free assay for glycosylphosphatidylinositol anchoring in African trypanosomes. Demonstration of a transamidation reaction mechanism.},
  author={Deepak Sharma and Jolanta Vidugirienė and James D Bangs and Anant K. Menon},
  journal={The Journal of biological chemistry},
  year={1999},
  volume={274 23},
  pages={
          16479-86
        }
}
We established an in vitro assay for the addition of glycosyl-phosphatidylinositol (GPI) anchors to proteins using procyclic trypanosomes engineered to express GPI-anchored variant surface glycoprotein (VSG). The assay is based on the premise that small nucleophiles, such as hydrazine, can substitute for the GPI moiety and effect displacement of the membrane anchor of a GPI-anchored protein or pro-protein causing release of the protein into the aqueous medium. Cell membranes containing pulse… CONTINUE READING

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