Despite the long history [Kaiser, E., 1956. Enzymatic activity of
spider venoms. In: Buckley, E.E., Porges, N. (Eds.),
Venoms. American Association for the Advancement of Science, Washington, DC, pp. 91-93] on proteolytic activity, no study so far claims the isolation of a
serine protease from the
spider venom/
venom gland extract. Therefore, the present study describes the isolation and characterization of a low molecular weight
serine protease from Hippasa agelenoides
venom gland extract. The
protease (Hag-
protease) was purified to homogeneity using the combination of gel-permeation and ion-exchange chromatography. The molecular mass was found to be 16.350 kDa by matrix-assisted
laser desorption ionization time of flight (MALDI-TOF) mass spectrometry. Hag-
protease was optimally active at pH 7.5 and temperature of 37 degrees C. PMSF abolished the
enzyme activity while
EDTA,
EGTA, IAA, 1, 10-phenanthrolene did not. It hydrolyzed
proteins such as
casein,
fibronectin and
collagen type-I dose dependently but did not degrade
gelatin and
collagen type-IV. The isolated
protease was non-lethal and devoid of hemorrhagic, myotoxic and
edema forming activities. The light microscopy of Hag-
protease treated skin tissue sections at the site of injection showed extensive damage of extracellular matrix (ECM) of hypodermis without causing any damage to blood vessels and capillaries. Similar damage of ECM of muscle tissue sections without affecting myocytes was noticed. Hag-
protease was found to be procoagulant in property when studied plasma recalcification time.