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Cloning and expression in Escherichia coli of a cDNA encoding a developmentally regulated Ca(2+)-binding protein from
B Coukell1, J Moniakis, A Grinberg
1Department of Biology, York University, Ont., Canada.
Abstract:
We have cloned a full-length cDNA from Dictyostelium discoideum which encodes a new Ca(2+)-binding protein. The deduced protein (termed CBP1) is composed of 156 amino acids and contains four consensus metal-ligating loop sequences found in helix-loop-helix motifs of many Ca(2+)-binding proteins. When expressed in bacteria as a GST fusion protein, CBP1 binds Ca2+ in a 45Ca2+ overlay assay. CBP1 exhibits little amino acid sequence homology with Dictyostelium calmodulin or calfumirin-1 (CAF-1) except in the putative Ca(2+)-binding regions. Moreover, unlike calmodulin and CAF-1 expression, CBP1 mRNA is expressed preferentially during the multicellular stages of development.