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

Yoko Furukawa, Hiroomi Tamura, Hiroh Ikezawa

研究成果: Article

16 引用 (Scopus)

抄録

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.

元の言語English
ページ(範囲)273-278
ページ数6
ジャーナルBBA - Biomembranes
1190
発行部数2
DOI
出版物ステータスPublished - 1994 3 23
外部発表Yes

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Glycosylphosphatidylinositols
5'-Nucleotidase
Liver
Amino Acids
COS Cells
Hydrophobicity
Hydrophobic and Hydrophilic Interactions
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology

これを引用

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title = "Mutational analysis of the COOH-terminal hydrophobic domain of bovine liver 5′-nucleotidase as a signal for glycosylphosphatidylinositol (GPI) anchor attachment",
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.",
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author = "Yoko Furukawa and Hiroomi Tamura and Hiroh Ikezawa",
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AU - Furukawa, Yoko

AU - Tamura, Hiroomi

AU - Ikezawa, Hiroh

PY - 1994/3/23

Y1 - 1994/3/23

N2 - 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.

AB - 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.

KW - 5′-Nucleotidase

KW - COS cell

KW - Glycosylphosphatidylinositol anchor

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