Mutational analysis of the COOH-terminal hydrophobic domain of bovine liver 5′-nucleotidase as a signal for glycosylphosphatidylinositol (GPI) anchor attachment

  • Furukawa Y
  • Tamura H
  • Ikezawa H
  • 3

    Readers

    Mendeley users who have this article in their library.
  • 16

    Citations

    Citations of this article.

Abstract

In order to address the minimum domain of the COOH-terminal hydrophobic region responsible for GPI modification of bovine liver 5′-nucleotidase, we constructed a series of the deletion mutants of the COOH-terminus and expressed them in COS cells. Cells transfected by the deletion mutant of 6 amino acids (-IIILYQ) from the hydrophobic domain (-FSLIFLSVLAVIIILYQ) did not show any elevation of cell surface-associated 5′-nucleotidase activity, whereas the 2 (-YQ) or 4 (-ILYQ) amino acid deletion mutant retained the bovine liver-derived activity on the cell surface as a GPI-anchored protein. Loss of half the hydrophobic domain (6 or 8 amino acids) resulted in accumulation of the activity in the cell. On the other hand, deletion of the whole hydrophobic domain (17 amino acids) or the entire cleaved-off domain (25 amino acids) made the product secreted into the medium. In conclusion, the hydrophobicity of 13 amino acids in length was enough for the GPI modification of the bovine liver 5′-nucleotidase. © 1994.

Author-supplied keywords

  • 5′-Nucleotidase
  • COS cell
  • Glycosylphosphatidylinositol anchor

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • Yoko Furukawa

  • Hiro omi Tamura

  • Hiroh Ikezawa

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free