Purification of a fragment obtained by autolysis of a PIIIb-SVMP from Bothrops alternatus venom

VAN DE VELDE, ANDREA CAROLINA; GAY, CLAUDIA CAROLINA; MORITZ, MILENE NÓBREGA DE OLIVEIRA; DOS SANTOS, PATTY KARINA; BUSTILLO, SOLEDAD; RODRÍGUEZ, JUAN PABLO; ACOSTA, OFELIA CRISTINA; BISCOGLIO, MIRTHA JOSEFA; SELISTRE-DE-ARAUJO, HELOISA SOBREIRO; LEIVA, LAURA CRISTINA

INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES

ELSEVIER SCIENCE BV

Año: 2018 vol. 113 p. 205 - 211

0141-8130

Snake Venom Metalloproteinases (SVMPs) represent 43.1% of the components in Bothrops alternatus venom and play an important role in envenomation. Disintegrins and disintegrin-like domains are released by proteolytic processing of PII and PIII classes of SVMPs respectively and are potent inhibitors of integrin?ligand interaction. Baltergin is a PIIIb-SVMP isolated from this venom and able to undergo autolysis in vitro, giving rise to a stable disintegrin-like/cystein-rich fragment (baltergin-DC). Conditions of baltergin autolysis were adjusted in order to carry out the purification of baltergin-DC and its effect on cell adhesion was studied. Autolysis was maximal at 37 °C and a pH range of 7.0?8.0. Baltergin-DC amino-terminal sequence begins with IISPPVCGNELLEVGEECDCGTPENCQNECCDAATC, which shows a high degree of homology with other disintegrin-like proteins. Baltergin and purified baltergin-DC were both able to inhibit C2C12 adhesion to fetal bovine serum (FBS) coated plates, indicating that a non-catalytic process is involved, probably mediated by binding to membrane integrins. Baltergin-DC, lacking proteolytic action, becomes an attractive molecule for future studies on blocking integrin?ligand interactions.