Structural and functional relationships in the virulence-associated cathepsin L proteases of the parasitic liver fluke, Fasciola hepatica - PubMed (original) (raw)

Comparison of the collagen cleaving activities of wild type FheCL1, variant FheCL1 L205A, variant FheCL1 L67Y, and wild type FheCL2. A, type I collagen was incubated with FheCL1 and FheCL2 at pH 4.0 and 5.5 and at 28 °C for 3 h, and the reaction was analyzed by 4–20% SDS-PAGE: lane 1, collagen alone; lane 2 collagen plus FheCL1 pH 4.0; lane 3, collagen plus FheCL2 pH 4.0; lane 4, collagen alone; lane 5, collagen plus FheCL1, pH 5.5; lane 6, collagen plus FheCL2, pH 5.5. B, type I collagen was incubated with active recombinant peptidase (5.47 μ

m

) at 28 °C at neutral pH (PBS, pH 7.3) for 20 h, and the reactions analyzed as above: lane 1, collagen alone (i.e. no peptidase added); lane 2, collagen plus FheCL1; lane 3, collagen plus FheCL1 L209A; lane 4, collagen plus FheCL1 L67Y; lane 5, collagen plus FheCL2. Molecular mass standards are indicated on the right, and collagen chains (_α_1, _α_2, β 11, β 12, and γ, see Ref. 44) are indicated on the left.