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

Yoko Furukawa, Hiro omi Tamura, Hiroh Ikezawa

Research output: Contribution to journalArticle

16 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)273-278
Number of pages6
JournalBBA - Biomembranes
Volume1190
Issue number2
DOIs
Publication statusPublished - 1994 Mar 23
Externally publishedYes

Keywords

  • 5′-Nucleotidase
  • COS cell
  • Glycosylphosphatidylinositol anchor

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

Fingerprint Dive into the research topics of 'Mutational analysis of the COOH-terminal hydrophobic domain of bovine liver 5′-nucleotidase as a signal for glycosylphosphatidylinositol (GPI) anchor attachment'. Together they form a unique fingerprint.

  • Cite this