Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9442974 | Experimental Parasitology | 2005 | 6 Pages |
Abstract
Serine proteases are one of the biologically most important and widely distributed enzyme families. A protease capable of degrading the substrate Suc-AAF-AMC was isolated from axenically grown trophozoites of Entamoeba histolytica. The enzyme was purified by ion-exchange chromatography and electroelution, and appeared on 2D-PAGE as a spot of 60 kDa and pI of 4.65. Data obtained from zymogram suggest the active protease is present either as homodimer (130 kDa) or homotetramer (250 kDa). The optimal temperature of the enzyme was 37 °C, and it exhibited activity over a broad pH range. The protease was strongly inhibited by TPCK and chelating agents. The enzymatic activity was restored upon addition of calcium. BLAST analysis with the sequence of internal peptides of the protein revealed two open reading frames within the genome of E. histolytica, homologous to members of the family S28, clan SC of serine proteases.
Keywords
TPCKdi-isopropyl fluorophosphatePRCPSBTIN-tosyl-l-phenylalanine chloromethyl ketoneTLCK2D-PAGEDPPE-64PMSFDFPEntamoeba histolyticaEDTAEthylenediaminetetraacetic acidtwo-dimensional polyacrylamide gel electrophoresisdipeptidyl peptidaseSerine proteasephenylmethylsulfonyl fluoridesoybean trypsin inhibitorProlylcarboxypeptidase
Related Topics
Life Sciences
Immunology and Microbiology
Parasitology
Authors
Minerva Paola Barrios-Ceballos, Norma Angélica MartÃnez-Gallardo, Fernando Anaya-Velázquez, David Mirelman, Felipe Padilla-Vaca,