In vitro kinase assays were performed with recombinant maltose-binding protein (MBP) tag-fused PfCDPK4 protein using synthetic peptides as substrates. Kinase reaction buffer contained CaCl2 or the calcium chelator EGTA. Triplicate reactions with either the CaCl2 buffer or the EGTA buffer were performed in parallel and the peptides were detected and quantified by LC-MS/MS. The suitability of a peptide as a substrate for PfCDPK4 was quantified as the depletion of the unmodified peptide as it was converted to phosphopeptide. Additionally, autophosphorylation of PfCDPK4 was assessed by performing the reaction in the absence of substrates followed by tryptic digest and LC-MS/MS of the resulting peptides to identify phosphorylation sites.
[doi:10.25345/C52V5N]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: plasmodium falciparum ; kinase ; phosphorylation
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Principal Investigators: (in alphabetical order) |
Kristian Swearingen, Institute for Systems Biology, United States |
| Submitting User: | kswearingen |
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