FEBS Letters · 1996 · 27 citations · 29 references
Life SciencesDevelopmental BiologyMolecular BiologyComparative GenomicsGenomicsPlant Molecular BiologyCalcium-binding ProteinsProtein Interaction DomainsProtein StructureAmino AcidsCalcium-calmodulin SignalingCalmodulin-mediated RegulationEf-hand MotifsEf-hand ProteinsIntron Consensus SequencesPolyclonal AntibodiesWestern Blot Analysis
A full-length genomic DNA fragment that codes for a novel EF-hand protein Dictyostelium discoideum was cloned and sequenced. The protein is composed of 168 amino acids and contains four consensus sequences that are typical for (Ca2+)-binding EF-hand domains. The protein sequence exhibits only minor similarities to other calmodulin-type proteins from Dictyostelium. The genomic DNA harbors two short introns; their positions suggest that the gene is unrelated to the EF-hand proteins from the calmodulin group. Northern blot analysis showed that the mRNA level was significantly increased during development. Polyclonal antibodies raised against the recombinant protein recognized in Western blots a protein of about 20 kDa. Like the mRNA, also the protein was more abundant in developing cells. Overlay experiments with 45Ca2+ indicated that the EF-hands in fact have (Ca2+)-binding activity. The recent description of CBP1, another calmodulin-type Dictyostelium protein that is upregulated during development [Coukell et al. (1995) FEBS Lett. 362, 342-346], suggests that D. discoideum contains a family of EF-hand proteins that have specific functions during distinct steps of development. We therefore designate the protein described in this report as CBP2.
29
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4
Ulrich K. Laemmli · Nature · 1970 · 251.6K citations
A comprehensive set of sequence analysis programs for the VAX
John Devereux, Paul Haeberli, Oliver Smithies · Nucleic Acids Research · 1984 · 14.4K citations · Full text
Current Protocols in Molecular Biology
Tim Hunt · Journal of Cell Science · 1989 · 969 citations