Physico-chemical properties of brain cathepsin H

Cathepsin H (EC 3.4.22.16) from cow brain, purified to approximately 1800-fold with approximately 26% activity yield, hydrolysed BANA, Leu-2-NNap, Arg-2-NNap, and Met-2-NNap maximally at pH 6.5, 6.8, 7.0 and 7.2, respectively. It was activated by sulphydryl compounds and EDTA while sulphydryl alkylators and blockers were found to inhibit the enzyme activity. Met-2-NNap was found to be the best substrate followed by Thr-2-NNap, His-2-NNap, Leu-2-NNap, Arg-2-NNap and Ala-2-NNap, respectively. The Km values for hydrolysis of various substrates viz., Met-2-NNap, Leu-2-NNap, Arg-2-NNap, Arg-NNapOMe, Thr-2-NNap, His-2-NNap, BANA, Arg-pNA and Lys-pNA were 0.128, 0.167, 0.169, 0.288, 0.428, 0.500, 0.667, 0.195 and 0.476 mM, respectively. The temperature optima for hydrolysis of BANA and Leu-2-NNap were approximately 45 degrees C and approximately 50 degrees C with activation energies of approximately 13.7 and approximately 11.0 kcal mole-1, respectively. The enzyme was fairly stable upto 50 degrees C and between pH 4.0-7.5.