Biochemical and functional properties of a thrombin-like enzyme isolated from Bothrops pauloensis snake venom.
In the present study, a thrombin-like enzyme named BpSP-I was isolated from Bothrops pauloensis snake venom and its biochemical, enzymatic and pharmacological characteristics were determined. BpSP-I is a glycoprotein that contains both N-linked carbohydrates and sialic acid in its structure, with M(r)=34,000 under reducing conditions and pI approximately 6.4. The N-terminal sequence of the enzyme (VIGGDECDINEHPFL) showed high similarity with other thrombin-like enzymes from snake venoms. BpSP-I showed high clotting activity upon bovine and human plasma and was inhibited by PMSF, benzamidine and leupeptin. Moreover, this enzyme showed stability when examined at different temperatures (-70 to 37 degrees C), pH values (3-9) or in the presence of divalent metal ions (Ca(2+), Mg(2+), Zn(2+) and Mn(2+)). BpSP-I showed high catalytic activity upon substrates, such as fibrinogen, TAME, S-2238 and S-2288. It also showed kallikrein-like activity, but was unable to act upon factor Xa and plasmin substrates. Indeed, the enzyme did not induce hemorrhage, myotoxicity or edema. Taken together, our data showed that BpSP-I is in fact a thrombin-like enzyme isoform isolated from Bothrops pauloensis snake venom.
Costa
Fábio L S
FL
Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil.
Rodrigues
Renata S
RS
Izidoro
Luiz F M
LF
Menaldo
Danilo L
DL
Hamaguchi
Amélia
A
Homsi-Brandeburgo
Maria I
MI
Fuly
André L
AL
Soares
Sandro G
SG
Selistre-de-Araújo
Heloisa S
HS
Barraviera
Benedito
B
Soares
Andreimar M
AM
Rodrigues
Veridiana M
VM
D000595:Amino Acid Sequence; D000818:Animals; D017837:Bothrops; D003435:Crotalid Venoms / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D013917:Thrombin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D003435:Crotalid Venoms; D013917:Thrombin