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 language | English (US) |
|---|---|
| Pages (from-to) | 71-78 |
| Number of pages | 8 |
| Journal | Cell |
| Volume | 39 |
| Issue number | 1 |
| DOIs | |
| State | Published - 1984 |
| Externally published | Yes |
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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 journal › Article
}
TY - JOUR
T1 - Characterization and purification of a soluble protein controlling Ca-channel activity in paramecium
AU - Haga, Nobuyuki
AU - Forte, Michael
AU - Ramanathan, Rajeev
AU - Hennessey, Todd
AU - Takahashi, Mihoko
AU - Kung, Ching
PY - 1984
Y1 - 1984
N2 - 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 (
AB - 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 (
UR - http://www.scopus.com/inward/record.url?scp=0021674199&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021674199&partnerID=8YFLogxK
U2 - 10.1016/0092-8674(84)90192-2
DO - 10.1016/0092-8674(84)90192-2
M3 - Article
C2 - 6091917
AN - SCOPUS:0021674199
VL - 39
SP - 71
EP - 78
JO - Cell
JF - Cell
SN - 0092-8674
IS - 1
ER -