The GTP-binding regulatory proteins of neuroblastoma × glioma, NG108-15, and glioma, C6, cells. Immunochemical evidence of a pertussis toxin substrate that is neither Ni nor No

Graeme Milligan, Peter Gierschik, Allen M. Spiegel, Werner A. Klee

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

Amounts of the guanine nucleotide binding regulatory proteins which are also pertussis toxin substrates (such as Ni and No) were measured in rat glioma, C6BU-1, cells and in neuroblastoma × glioma, NG108-15, hybrid cells. Measurements were performed both by quantitating pertussis toxin catalyzed ADP-ribosylation and by quantitative immunoblotting with affinity purified antibodies specific for Ni or No. The amounts of pertussis toxin substrate in C6 and NG108-15 cells are 7.5 and 0.6 pmol mg membrane protein, respectively. These levels are minimum values and higher estimates of the total amounts of N proteins in the two cells are obtained by quantitative immunoblot analysis of the β-subunit common to all N proteins. Immunoblots with specific antibodies show that NG108-15 cells contain 3.8 pmol mg of No and detectable but small ( < 0.1 pmol mg) amounts of Ni. In contrast, C6 cell membranes contain no detectable No and only 0.14 pmol mg Ni. Thus, C6 cells contain large amounts of a pertussis toxin substrate which is neither Ni nor No.

Original languageEnglish (US)
Pages (from-to)225-230
Number of pages6
JournalFEBS Letters
Volume195
Issue number1-2
DOIs
StatePublished - Jan 20 1986
Externally publishedYes

Keywords

  • GTP-binding protein Immunoblotting Pertussis toxin ADP-ribosylation

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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