Inhibitory properties of the anti-bothropic complex from Didelphis albiventris serum on toxic and pharmacological actions of metalloproteases and myotoxins from Bothrops asper venom.
Anti-bothropic complex (ABC) was isolated from the serum of the South American opossum (Didelphis albiventris) by single-step affinity chromatography using a Sepharose-immobilized metalloprotease (BaP1) from Bothrops asper as the binding protein. Biochemical characterization of ABC showed the presence of two glycosylated subunits of 43 and 45 kDa, respectively, with an isoelectric point < 4. The two subunits were separated by ion-exchange HPLC. The N-terminal sequences of both subunits (LKAMDPTPXLWIETESP, where X is Arg-9 and Pro-9, respectively) showed a high degree of identity with other serum inhibitors isolated from different marsupials. Functional studies pointed out that ABC inhibits the hemorrhagic and proteolytic activities on fibrin, fibrinogen, and casein induced by the metalloproteases BaP1 and BaH4 isolated from B. asper venom. In addition to the anti-hemorrhagic and anti-proteolytic activities, ABC also showed anti-myotoxic, anti-lethal, and anti-edematogenic effects against myotoxic phospholipases A(2) isolated from the same venom. Moreover, it had inhibitory effects on the phospholipase A(2) activity of the crude venom as well as the isolated venom phospholipases A(2).
Trento
E P
EP
Departamento de Bioquímica, UNIC, Cuiabá-MT, Brazil.
Garcia
O S
OS
Rucavado
A
A
França
S C
SC
Batalini
C
C
Arantes
E C
EC
Giglio
J R
JR
Soares
A M
AM
D000596:Amino Acids / Q000737:chemistry; D000818:Animals; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D017837:Bothrops; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D003435:Crotalid Venoms / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000201:enzymology; D054501:Group II Phospholipases A2; D006470:Hemorrhage / Q000517:prevention & control; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors*; D009498:Neurotoxins / Q000037:antagonists & inhibitors*; D009893:Opossums / Q000097:blood*; D010741:Phospholipases A / Q000037:antagonists & inhibitors*; D030162:Reptilian Proteins; D020539:Sequence Analysis, Protein
D000596:Amino Acids; D001798:Blood Proteins; D002352:Carrier Proteins; D003435:Crotalid Venoms; D009498:Neurotoxins; D030162:Reptilian Proteins; C442303:anti-bothropic complex, Didelphis; D010741:Phospholipases A; D054501:Group II Phospholipases A2; C061056:myotoxin II, Bothrops asper; D008666:Metalloendopeptidases