Heike Dammann's research while affiliated with Institut Pasteur and other places
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Publications (8)
The catalytic subunit of the cAMP-dependent protein kinase (PKA) from Dictyostelium discoideum contains several domains, including an unusually long N-terminal extension preceding a highly conserved catalytic core. We transformed the aggregationless PkaC-null strain with several deletion constructs of both domains. Strains transformed with genes ex...
The catalytic subunit of Ca2+/calmodulin-dependent protein phosphatase (calcineurin A) was overexpressed about 50-fold in Dictyostelium discoideum cells transformed with a vector containing the cDNA for D. discoideum calcineurin A under control of the actin-6 promoter. In crude lysates from the overexpressing cell line, high Ca2+/calmodulin-stimula...
cDNA clones for the catalytic subunit of Ca2+/calmodulin(CaM)-dependent protein phosphatase (calcineurin A, protein phosphatase 2B) from Dictyostelium discoideum were isolated by functional screening of a lambda gt11 lysogen expression library with labeled Dictyostelium CaM. A complete cDNA of 2146 bp predicts a protein of 623 amino acids with homo...
Nucleoside-diphosphate kinase (EC 2.7.4.6) catalyzes phosphate exchange between nucleoside triphosphates and nucleoside diphosphates. Its 17 kDa subunits are highly conserved throughout evolution in both sequence and tertiary structure. Using site-directed mutagenesis we investigated the function of 8 amino acids (Lys16, Tyr56, Arg92, Thr98, Arg109...
We have initiated a systematic study of Ca2+/calmodulin-regulated enzymes in the cellular slime mold Dictyostelium discoideum. Using 125I-labelled D. discoideum calmodulin (CaM) as a functional probe, several Ca2+/CaM-binding proteins were detected in crude cell lysates. Proteins with apparent molecular weights of 22 kDa and 78-80 kDa, respectively...
Isolation of cDNA clones from lambda gt11 phage libraries by functional screening is limited by the low amount of lacZ-cDNA-encoded fusion protein synthesized in an isolated phage plaque. The amount of specific cDNA-encoded protein can be significantly enhanced by expression in bacterial colonies rather than phage plaques. Escherichia coli was lyso...
Citations
... Control cells were transformed with empty pDXA-3H vector. The pkaC cDNA was either expressed from its endogenous promoter (plasmid p188) (1) or under the control of the constitutively active act6 promoter (plasmid p332) (12). ...
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... EGTA together with A23187 have been shown to decrease intracellular Ca 2 þ levels in D. discoideum cells by more than 2 fold (Baskar et al., 2000). W-7 is a specific antagonist of CaM that antagonizes CaM binding to its effectors in D. discoideum (Hidaka et al., 1981;Lydan and O'Day, 1988b;Winckler et al., 1991;Myre and O'Day, 2004). ...
... Probing protein blots from Dictyostelium cell extracts with '27-labeled Dictyostelium CaM revealed several specific high-affinity CaMbinding proteins . In parallel, we have developed a technique for direct, functional isolation of cDNA clones for CaM-binding proteins by probing cDNA expression libraries with labeled CaM (Mutzel et al., 1990). ...
... NDPK-B belongs to the family of NDPKs that were discovered around the 1950s in yeast and pigeon breast muscle [163,164]. NDPKs are housekeeping enzymes that are primarily involved in the catalytic transfer of γ-phosphate between a nucleoside 5'-triphosphate (NTP) and a nucleoside 5'-diphosphate (NDP) via a ping-pong mechanism which involves the formation of a high energy phosphate intermediate, e.g. on Histidine118 of NDPK-B [165,166]. ...
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... Historically, CaMBPs in Dictyostelium have been revealed through directed studies aimed at confirming whether a suspected protein binds CaM [16][17][18][19][20][21][22][23][24][25][26]. In addition, the CaM-binding overlay technique (CaMBOT), which involves separating proteins by SDS-PAGE and then performing a gel overlay with recombinant radiolabelled CaM (35[S]-CaM), has been useful for identifying putative CaMBPs in a biological sample [27]. ...
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... CaM is an activator of CaN, not only in higher eukaryotes but also in ciliates (Kissmehl et al. 1997) and in myxamoebae (Aichem and Mutzel 2001;Hellstern et al. 1997). Already in protozoa, the CaN-A subunit contains a CaMbinding domain. ...
