Comparative structural analysis of calmodulins from Trypanosoma brucei, T. congolense, T. vivax, Tetrahymena thermophila and bovine brain.
Calmodulin is an intracellular calcium receptor protein utilized extensively by eukaryotic cells to mediate responsiveness to calcium signals. The present study evaluates the effects on protein structure of amino acid substitutions in trypanosome calmodulin. Calmodulin conformation, hydrophobicity and antigenic determinants are compared among Trypanosoma brucei, Trypanosoma congolense, Trypanosoma vivax, Tetrahymena thermophila and bovine brain. Trypanosome calmodulin differs from brain and Tetrahymena calmodulins based upon isoelectric point, retention time on a C-2/C-18 reverse phase column and interaction with polyclonal antibodies against trypanosome calmodulin by radioimmunoassay or Western procedures. These same analyses do not distinguish trypanosome calmodulins from each other. Polyclonal antibodies against Tetrahymena calmodulin are equally specific and do not recognize the trypanosome or brain calmodulins. Calcium-induced exposure of hydrophobic binding sites are quantitated using the fluorescent probe, N-phenyl-1-naphthylamine. All calmodulins, regardless of source, enhance the fluorescence of N-phenyl-1-naphthylamine 3-4 fold in the presence of calcium. These data demonstrate the extent to which functional calmodulins vary in their structures. We conclude that African trypanosomes share a common calmodulin that is structurally distinct from calmodulin of vertebrates or Tetrahymena.
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D000818:Animals; D000906:Antibodies; D001923:Brain Chemistry; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000032:analysis* / Q000145:classification / Q000276:immunology; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D000939:Epitopes / Q000032:analysis; D007526:Isoelectric Point; D011487:Protein Conformation; D011817:Rabbits; D013045:Species Specificity; D013768:Tetrahymena / Q000032:analysis* / Q000145:classification; D014345:Trypanosoma / Q000032:analysis* / Q000145:classification; D014346:Trypanosoma brucei brucei / Q000032:analysis / Q000145:classification; D014348:Trypanosoma congolense / Q000032:analysis / Q000145:classification
D000906:Antibodies; D002147:Calmodulin; D000939:Epitopes; D002118:Calcium