Bodo sp., a free-living flagellate, expresses divergent proteolytic activities from the closely related parasitic trypanosomatids.
We report the characterization of cell-associated and extracellular peptidases of Bodo sp., a free-living flagellate of the Bodonidae family, order Kinetoplastida, which is considered ancestral to the trypanosomatids. This bodonid isolate is phylogenetically related to Bodo caudatus and Bodo curvifilus. The proteolytic activity profiles of Bodo sp. were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis containing co-polymerized gelatin, casein, hemoglobin, or bovine serum albumin as substrates. The enzymatic complex degraded gelatin better in acidic pH, and under these conditions four proteolytic bands (120, 100, 90, and 75 kDa) were detected in the cellular or extracellular extracts. Two peptidases (250 and 200 kDa) were exclusively detected with the substrate casein. All these enzymes belong to the serine peptidase class, based on inhibition by aprotinin and phenylmethylsulfonyl fluoride. This is the first biochemical characterization of peptidases in a free-living Bodo sp., potentially providing insight into the physiology of these protozoa and the evolutionary importance of peptidases to the order Kinetoplastida as some of these enzymes are important virulence factors in pathogenic trypanosomatids.
D'Avila-Levy
Claudia M
CM
Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, Brazil. davila.levy@ioc.fiocruz.br
Volotão
Aline C C
AC
Araújo
Fernanda M
FM
de Jesus
José B
JB
Motta
Maria Cristina M
MC
Vermelho
Alane B
AB
Santos
André L S
AL
Branquinha
Marta H
MH
D000818:Animals; D007611:Aprotinin / Q000494:pharmacology; D016000:Cluster Analysis; D003059:Cocos / Q000469:parasitology; D016054:DNA, Protozoan / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology; D020459:Genes, rRNA; D016830:Kinetoplastida / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010664:Phenylmethylsulfonyl Fluoride / Q000494:pharmacology; D010802:Phylogeny; D015800:Protozoan Proteins / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*; D016053:RNA, Protozoan / Q000235:genetics; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012697:Serine Endopeptidases / Q000032:analysis* / Q000737:chemistry / Q000235:genetics*
D016054:DNA, Protozoan; D004275:DNA, Ribosomal; D004791:Enzyme Inhibitors; D015800:Protozoan Proteins; D016053:RNA, Protozoan; D012337:RNA, Ribosomal, 18S; D010664:Phenylmethylsulfonyl Fluoride; D007611:Aprotinin; D012697:Serine Endopeptidases