Publication Type

Journal Article

Journal Name

Insect Biochemistry and Molecular Biology

Publication Date

1-1-1992

Abstract

Two proteinases designated 1 and 2, purified from the gut of partially engorged female, Rhipicephalus appendiculatus have been characterized. Both proteinases hydrolysed denatured haemoglobin (Hb) at acid pH, with Michaelis constants (KM) of 1.19 g/100 ml (proteinase 1) and 0.71 g/100 ml (proteinase 2). Pepstatin competitively inhibited both proteinases, with a K1 of 1.7 nM (proteinase 2). Both proteinases were also inhibited by 1 mM phenylmethylsulphonyl fluoride (PMSF). The pH optimum for proteinase 1 was 3.0, with a smaller but distinct second peak at pH 4.6. For proteinase 2, peak activity was at pH 2.6 with a second smaller peak at pH 5.0. Both enzymes rapidly lost activity at 50°C and above. Thermal inactivation and inhibition by pepstatin were pH-dependent for proteinase 2 but not for proteinase 1. On the basis of these results, proteinases 1 and 2 have been classified as aspartic proteinases. © 1992.

Keywords

aspartic proteinases, cathepsin D, digestive enzymes, Rhipicephalus appendiculatus

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