A hyaluronidase from Tityus serrulatus scorpion venom: isolation, characterization and inhibition by flavonoids

• Published in: Toxicon (Oxford) Vol. 39; no. 10; pp. 1495 - 1504
• Main Authors: Pessini, Andréa C, Takao, Tânia T, Cavalheiro, Elisângela C, Vichnewski, Walter, Sampaio, Suely V, Giglio, José R, Arantes, Eliane C
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01-10-2001; Elsevier Science
• Subjects: Adjuvants, Immunologic - metabolism; Animal poisons toxicology. Antivenoms; Animals; Aspartate Aminotransferases - blood; Biological and medical sciences; Buthidae; Chromatography, Ion Exchange; Creatine Kinase - blood; Electrophoresis, Polyacrylamide Gel; Flavonoid; flavonoids; Flavonoids - metabolism; Hot Temperature; Hyaluronic Acid - metabolism; Hyaluronidase; Hyaluronoglucosaminidase - analysis; Hyaluronoglucosaminidase - antagonists & inhibitors; Hyaluronoglucosaminidase - isolation & purification; Hyaluronoglucosaminidase - metabolism; Hydrogen-Ion Concentration; Insect Proteins; Kinetics; L-Lactate Dehydrogenase - blood; Male; Medical sciences; Molecular Weight; Neurotoxins - toxicity; Rats; Rats, Wistar; Scorpion venom; Scorpion Venoms - chemistry; Scorpion Venoms - toxicity; Scorpiones; Scorpions; Serum enzymes; Sesquiterpenes - metabolism; Structure-Activity Relationship; T. serrulatus; Tityus serrulatus; Toxicology; Hyaluronidase; T. serrulatus; Flavonoid; Scorpion venom; Serum enzymes; Characterization; Toxin; Arachnida; Scorpionides; Purification; Arthropoda; Venom; Animal origin; Isolation; Invertebrata
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/S0041-0101(01)00122-2

The purification procedure of a hyaluronidase from
Tityus serrulatus scorpion venom is described. It involves basically an ion-exchange chromatography on CM-cellulose at pH 7.8 followed by a rechromatography of the active fraction on the same column at pH 4.7. The optima pH and temperature for maximum activity of the isolated enzyme was 6.0 and 40°C, respectively. Its
K
M was 69.7
μg/ml at 37°C and its specific activity was 19,900±1,730 turbidity reducing units (TRU)/mg against 845±88
TRU/mg for the whole desiccated venom, representing a 23- to 24-fold purification range. The hyaluronidase activity of the purified protein (51
kDa) was inhibited by some flavonoid compounds. This article also showed that
T. serrulatus hyaluronidase affected on the activity of the venom's major toxin, tityustoxin-I (TsTX-I or Ts1), as reflected by alterations in the serum levels of creatine kinase (CK), lactate dehydrogenase (LD) and aspartate aminotransferase (AST) following injection of TsTX-I, in the presence or absence of hyaluronidase.