Structural and functional properties of calmodulin from the eukaryotic microorganism Dictyostelium discoideum.
Calmodulin was purified from the eukaryotic microorganism Dictyostelium discoideum and characterized in terms of its nearly complete primary structure and quantitative activator activity. The strategy for amino acid sequence analysis took advantage of the highly conserved structure of calmodulin and employed a new procedure for limited cleavage of calmodulin that uses a protease from mouse submaxillary gland. Fourteen amino acid sequence differences between Dictyostelium and bovine calmodulin were identified unequivocally, as well as an unmethylated lysine at residue 115 instead of N epsilon, N epsilon, N epsilon-trimethyllysine. Seven of the amino acid substitutions in Dictyostelium calmodulin are novel in that the residues at these positions are invariant in all calmodulin sequences previously examined, most notably an additional residue at the carboxy terminus. Comparison of the Dictyostelium calmodulin sequence with other calmodulin sequences shows that the region with the greatest extended sequence identity includes parts of the first and second structural domains and the interdomain region between domains 1 and 2. Dictyostelium calmodulin activated bovine brain cyclic nucleotide phosphodiesterase in a manner indistinguishable from that of bovine brain calmodulin. However, Dictyostelium calmodulin activated pea NAD kinase to a maximal level 4.6-fold greater than that produced by bovine brain calmodulin. This functional difference demonstrates the potential biological importance of the limited number of amino acid sequence differences between Dictyostelium calmodulin and other calmodulins and provides further insight into the structure, function, and evolution of the calmodulin family of proteins.
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology; D002147:Calmodulin / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D002417:Cattle; D003488:Cyanogen Bromide; D004023:Dictyostelium / Q000378:metabolism*; D004789:Enzyme Activation; D051379:Mice; D010446:Peptide Fragments; D010447:Peptide Hydrolases; D013363:Submandibular Gland / Q000201:enzymology; D013820:Thermolysin; D014357:Trypsin
D002147:Calmodulin; D010446:Peptide Fragments; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D010447:Peptide Hydrolases; D014357:Trypsin; D013820:Thermolysin; D003488:Cyanogen Bromide