Characterization and purification of a soluble protein controlling Ca-channel activity in paramecium

Nobuyuki Haga, Michael Forte, Rajeev Ramanathan, Todd Hennessey, Mihoko Takahashi, Ching Kung

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

The analysis of the voltage-sensitive Ca++ channel of the unicellular eucaryote, Paramecium has been extended to a biochemical level based on recent observations that the transfer of cytoplasm from wild-type cells into mutants lacking Ca++-channel function ("pawn" in P. tetraurelia and "CNR" in P. caudatum) causes mutant cells to regain Ca++-channel function. We have microinjected various cytoplasmic fractions into mutant cells and measured the restored Ca++-channel function using a convenient behavioral assay. Following the "curing" activity, we characterized and purified the component from wild-type cytoplasm that can restore the function missing in cells carrying mutations in the cnrC gene. The curing factor is not an RNA, but a heat-labile, -SH-containing protein that appears to affect existing mutant channels on the ciliary membrane. We have purified this factor over 500-fold from the soluble cytoplasm using conventional techniques. The protein is of low apparent molecular weight (

Original languageEnglish (US)
Pages (from-to)71-78
Number of pages8
JournalCell
Volume39
Issue number1
DOIs
StatePublished - 1984
Externally publishedYes

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Paramecium
Purification
Cytoplasm
Curing
Proteins
Urodela
Regain
Assays
Hot Temperature
Genes
Molecular Weight
Molecular weight
RNA
Membranes
Mutation
Electric potential

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Characterization and purification of a soluble protein controlling Ca-channel activity in paramecium. / Haga, Nobuyuki; Forte, Michael; Ramanathan, Rajeev; Hennessey, Todd; Takahashi, Mihoko; Kung, Ching.

In: Cell, Vol. 39, No. 1, 1984, p. 71-78.

Research output: Contribution to journalArticle

Haga, Nobuyuki ; Forte, Michael ; Ramanathan, Rajeev ; Hennessey, Todd ; Takahashi, Mihoko ; Kung, Ching. / Characterization and purification of a soluble protein controlling Ca-channel activity in paramecium. In: Cell. 1984 ; Vol. 39, No. 1. pp. 71-78.
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