ArticleThe Biochemical journal1995
Mutation of an EF-hand Ca(2+)-binding motif in phospholipase C of Dictyostelium discoideum: inhibition of activity but no effect on Ca(2+)-dependence.
Article in The Biochemical journal, 1995. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
11 citing papers in PubMed, 24 citations in OpenAlex.
- Article
- Article
- Ca2+ Regulation of Trypanosoma brucei Phosphoinositide Phospholipase C.Eukaryotic cell · 2015Article
- Four key signaling pathways mediating chemotaxis in Dictyostelium discoideum.The Journal of cell biology · 2008Article
- Phospholipase C regulation of phosphatidylinositol 3,4,5-trisphosphate-mediated chemotaxis.Molecular biology of the cell · 2007Article
- Essential role of PI3-kinase and phospholipase A2 in Dictyostelium discoideum chemotaxis.The Journal of cell biology · 2007Article
- Chemoattractants and chemorepellents act by inducing opposite polarity in phospholipase C and PI3-kinase signaling.The Journal of cell biology · 2007Article
- Spectroscopic characterization of the EF-hand domain of phospholipase C delta1: identification of a lipid interacting domain.Archives of biochemistry and biophysics · 2005Article
- The distinctive functions of the two structural calcium atoms in bovine pancreatic deoxyribonuclease.Protein science : a publication of the Protein Society · 2002Article
- Phospholipase Cdelta regulates germination of Dictyostelium spores.BMC cell biology · 2001Article
- Molecular and enzymatic characterization of three phosphoinositide-specific phospholipase C isoforms from potato.Plant physiology · 1998Article
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Phosphoinositide-specific phospholipase C (PLC) is dependent on Ca2+ ions for substrate hydrolysis. The role of an EF-hand Ca(2+)-binding motif in Ca(2+)-dependent PLC activity was investigated by site-directed mutagenesis of the Dictyostelium discoideum PLC enzyme. Amino acid residues with oxygen-containing side chains at co-ordinates x, y, z, -x and -z of the putative Ca(2+)-binding-loop sequence were replaced by isoleucine (x), valine (y) or alanine (z, -x and -z). The mutated proteins were expressed in a Dictyostelium cell line with a disrupted plc gene displaying no endogenous PLC activity, and PLC activity was measured in cell lysates at different Ca2+ concentrations. Replacement of aspartate at position x, which is considered to play an essential role in Ca2+ binding, had little effect on Ca2+ affinity and maximal enzyme activity. A mutant with substitutions at both aspartate residues in position x and y also showed no decrease in Ca2+ affinity, whereas the maximal PLC activity was reduced by 60%. Introduction of additional mutations in the EF-hand revealed that the Ca2+ concentration giving half-maximal activity was unaltered, but PLC activity levels at saturating Ca2+ concentrations were markedly decreased. The results demonstrate that, although the EF-hand domain is required for enzyme activity, it is not the site that regulates the Ca(2+)-dependence of the PLC reaction.
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