Direkt zum Inhalt
Merck
  • Molecular and biochemical characterization of a cathepsin B-like protease family unique to Trypanosoma congolense.

Molecular and biochemical characterization of a cathepsin B-like protease family unique to Trypanosoma congolense.

Eukaryotic cell (2008-02-19)
Carlos Mendoza-Palomares, Nicolas Biteau, Christiane Giroud, Virginie Coustou, Theresa Coetzer, Edith Authié, Alain Boulangé, Théo Baltz
ZUSAMMENFASSUNG

Cysteine proteases have been shown to be essential virulence factors and drug targets in trypanosomatids and an attractive antidisease vaccine candidate for Trypanosoma congolense. Here, we describe an important amplification of genes encoding cathepsin B-like proteases unique to T. congolense. More than 13 different genes were identified, whereas only one or two highly homologous genes have been identified in other trypanosomatids. These proteases grouped into three evolutionary clusters: TcoCBc1 to TcoCBc5 and TcoCBc6, which possess the classical catalytic triad (Cys, His, and Asn), and TcoCBs7 to TcoCBs13, which contains an unusual catalytic site (Ser, Xaa, and Asn). Expression profiles showed that members of the TcoCBc1 to TcoCBc5 and the TcoCBs7 to TcoCBs13 groups are expressed mainly in bloodstream forms and localize in the lysosomal compartment. The expression of recombinant representatives of each group (TcoCB1, TcoCB6, and TcoCB12) as proenzymes showed that TcoCBc1 and TcoCBc6 are able to autocatalyze their maturation 21 and 31 residues, respectively, upstream of the predicted start of the catalytic domain. Both displayed a carboxydipeptidase function, while only TcoCBc1 behaved as an endopeptidase. TcoCBc1 exhibited biochemical differences regarding inhibitor sensitivity compared to that of other cathepsin B-like proteases. Recombinant pro-TcoCBs12 did not automature in vitro, and the pepsin-matured enzyme was inactive in tests with cathepsin B fluorogenic substrates. In vivo inhibition studies using CA074Me (a cell-permeable cathepsin B-specific inhibitor) demonstrated that TcoCB are involved in lysosomal protein degradation essential for survival in bloodstream form. Furthermore, TcoCBc1 elicited an important immune response in experimentally infected cattle. We propose this family of proteins as a potential therapeutic target and as a plausible antigen for T. congolense diagnosis.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, non-essential amino acids and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with sodium bicarbonate, without L-glutamine, ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Hämin, from bovine, ≥90%
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with ribonucleosides, deoxyribonucleosides and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Hämin, BioXtra, from Porcine, ≥96.0% (HPLC)
Sigma-Aldrich
IgG aus Rinderserum, reagent grade, ≥95% (SDS-PAGE), essentially salt-free, lyophilized powder
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, L-glutamine, and non-essential amino acids, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, with Earle′s salts and non-essential amino acids, without L-glutamine, phenol red and sodium bicarbonate, Modified, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts, stable glutamine, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Earle′s salts and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, HEPES Modification, with Earle′s salts, 25 mM HEPES and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with L-glutamine, ribonucleosides and deoxyribonucleosides, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, 10 ×, With Earle′s salts, without L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, with L-glutamine and sodium pyruvate, without ribonucleosides, deoxyribonucleosides and sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With stable glutamine and sodium bicarbonate, without ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With stable glutamine and sodium bicarbonate, with ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts, L-glutamine and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Modified, with Earle′s salts and reduced NaHCO3. without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Joklik Modification, with L-glutamine, without calcium chloride and sodium bicarbonate, suitable for cell culture
SAFC
Minimum Essential Medium Eagle, AutoMod, with Earle′s salts, without L-glutamine and sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Alpha Modification, With sodium bicarbonate and L-glutamine, without ribonucleosides and deoxyribonucleosides, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Joklik Modification, With sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, Spinner Modification, with Earle′s salts and sodium bicarbonate, without calcium chloride and L-glutamine, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
IgG aus Rinderserum, technical grade, ≥80% (SDS-PAGE), buffered aqueous solution
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts, L-glutamine and non-essential amino acids, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Minimum Essential Medium Eagle, With Hanks′ salts and sodium bicarbonate, without L-glutamine, liquid, sterile-filtered, suitable for cell culture
SAFC
Minimum Essential Medium Eagle, HEPES Modification, with Earle′s salts, L-glutamine and 25 mM HEPES, without sodium bicarbonate, powder, suitable for cell culture