Trypanosoma cruzi adenylyl cyclase is encoded by a complex multigene family.
The parasitic protozoan Trypanosoma cruzi undergoes several differentiation events during its life cycle. Some of these transitions are thought to involve activation of adenylyl cyclase via the binding of peptide ligands to the cell surface. Here we describe the characterisation of the adenylyl cyclase gene family of T. cruzi. Two complete genes and one pseudogene have been sequenced. The protein products appear to have a large extracellular domain, a single transmembrane helix and a cytosolic catalytic domain. The adenylyl cyclase genes are present on at least six chromosomes and are scattered rather than clustered. They form a large polymorphic family in which the extracellular domain is particularly variable. An Escherichia coli adenylyl cyclase mutant could be complemented by expression of the catalytic domain of the T. cruzi enzyme. The recombinant protein had adenylyl cyclase activity in vitro, which was enhanced by increasing concentrations of divalent cations (Mn2+ > Mg2+). This constitutively active recombinant protein will be a useful tool for dissecting the catalytic mechanism of adenylyl cyclase.
Taylor
M C
MC
Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, UK. m.c.taylor@lshtm.ac.uk
Muhia
D K
DK
Baker
D A
DA
Mondragon
A
A
Schaap
P B
PB
Kelly
J M
JM
D000262:Adenylyl Cyclases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D020134:Catalytic Domain; D002874:Chromosome Mapping; D018076:DNA, Complementary; D017125:Genes, Protozoan; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D005810:Multigene Family; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014349:Trypanosoma cruzi / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development
D018076:DNA, Complementary; D011994:Recombinant Proteins; D000262:Adenylyl Cyclases