Identification, molecular and functional characterization of calmodulin gene of Phytomonas serpens 15T that shares high similarity with its pathogenic counterparts Trypanosoma cruzi

Protein J. 2011 Mar;30(3):212-9. doi: 10.1007/s10930-011-9322-4.

Abstract

In trypanosomatids, Ca²+-binding proteins can affect parasite growth, differentiation and invasion. Due to their importance for parasite maintenance, they become an attractive target for drug discovery and design. Phytomonas serpens 15T is a non-human pathogenic trypanosomatid that expresses important protein homologs of human pathogenic trypanosomatids. In this study, the coding sequence of calmodulin, a Ca²+-binding protein, of P. serpens 15T was cloned and characterized. The encoded polypeptide (CaMP) displayed high amino acid identity to homolog protein of Trypanosoma cruzi and four helix-loop-helix motifs were found. CaMP sequence analysis showed 20 amino acid substitutions compared to its mammalian counterparts. This gene is located on a chromosomal band with estimated size of 1,300 kb and two transcripts were detected by Northern blot analysis. A polyclonal antiserum raised against the recombinant protein recognized a polypeptide with an estimated size of 17 kDa in log-phase promastigote extracts. The recombinant CaMP retains its Ca²+-binding capacity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Amino Acid Sequence
  • Calcium / metabolism
  • Calmodulin / chemistry*
  • Calmodulin / genetics
  • Calmodulin / metabolism*
  • Cloning, Molecular*
  • Humans
  • Molecular Sequence Data
  • Protein Binding
  • Protein Structure, Secondary
  • Protozoan Proteins / chemistry*
  • Protozoan Proteins / genetics
  • Protozoan Proteins / metabolism*
  • Sequence Alignment
  • Trypanosoma cruzi / chemistry
  • Trypanosoma cruzi / genetics
  • Trypanosoma cruzi / metabolism
  • Trypanosomatina / chemistry
  • Trypanosomatina / genetics*
  • Trypanosomatina / metabolism

Substances

  • Calmodulin
  • Protozoan Proteins
  • Calcium

Associated data

  • GENBANK/DQ503481