Trypanosoma cruzi calmodulin: Cloning, expression and characterization

• Published in: Experimental parasitology Vol. 123; no. 4; pp. 326 - 333
• Main Authors: Garcia-Marchan, Yael, Sojo, Felipe, Rodriguez, Evelyn, Zerpa, Noraida, Malave, Caridad, Galindo-Castro, Ivan, Salerno, Milena, Benaim, Gustavo
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.12.2009; Elsevier
• Subjects: Animals; Antibodies, Protozoan - biosynthesis; Antibody; Biological and medical sciences; Calcium; Calcium-Transporting ATPases - blood; Calmodulin; Calmodulin - biosynthesis; Calmodulin - chemistry; Calmodulin - genetics; Calmodulin - immunology; Chickens; Circular Dichroism; Cloning; Cloning, Molecular; Electrophoresis, Polyacrylamide Gel; Enzyme Activation; Enzyme-Linked Immunosorbent Assay; Erythrocyte Membrane - enzymology; Female; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; Humans; Immunoglobulins - biosynthesis; Life cycle. Host-agent relationship. Pathogenesis; Protozoa; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Sequence Alignment; Sequence Analysis, DNA; Trypanosoma cruzi; Trypanosoma cruzi - chemistry; Trypanosoma cruzi - genetics; Trypanosoma cruzi - metabolism; Trypanosomatids; Trypanosomatids; Calcium; Antibody; Cloning; Calmodulin; Trypanosoma cruzi; Kinetoplastida; Protozoa; Trypanosomatidae; Parasite
• ISSN: 0014-4894; 1090-2449
• DOI: 10.1016/j.exppara.2009.08.010

We have cloned and expressed calmodulin (CaM) from Trypanosoma cruzi, for the first time, to obtain large amounts of protein. CaM is a very well conserved protein throughout evolution, sharing 100% amino acid sequence identity between different vertebrates and 99% between trypanosomatids. However, there is 89% amino acid sequence identity between T. cruzi and vertebrate CaMs. The results demonstrate significant differences between calmodulin from T. cruzi and mammals. First, a polyclonal antibody developed in an egg-yolk system to the T. cruzi CaM recognizes the autologous CaM but not the CaM from rat. Second, it undergoes a larger increase in the α-helix content upon binding with Ca 2+, when compared to CaM from vertebrates. Finally, two classic CaM antagonists, calmidazolium and trifluoperazine, capable of inhibiting the action of CaM in mammals when assayed on the plasma membrane Ca 2+ pump, showed a significant loss of activity when assayed upon stimulation with the T. cruzi CaM.