New insights into the structural elements involved in the skin haemorrhage induced by snake venom metalloproteinases.
Haemorrhage induced by snake venom metalloproteinases (SVMPs) is a complex phenomenon resulting in capillary disruption and extravasation. This study analysed structural elements important for the interaction of four Bothrops jararaca SVMPs of different domain organisation and glycosylation levels with plasma and extracellular matrix proteins: HF3 (P-III class) is highly glycosylated and ~80 times more haemorrhagic than bothropasin (P-III class), which has a minor carbohydrate moiety; BJ-PI (P-I class) is not haemorrhagic and the DC protein is composed of disintegrin-like/cysteine-rich domains of bothropasin. HF3, bothropasin and BJ-PI showed different degradation profiles of fibrinogen, fibronectin, vitronectin, von Willebrand factor, collagens IV and VI, laminin and Matrigel; however, only bothropasin degraded collagen I. In solid-phase binding assays HF3 and bothropasin interacted with fibrinogen, fibronectin, laminin, collagens I and VI; the DC protein bound only to collagens I and VI; however, no binding of BJ-PI to these proteins was detected. N-deglycosylation caused loss of structural stability of bothropasin and BJ-PI but HF3 remained intact, although its haemorrhagic and fibrinogenolytic activities were partially impaired. Nevertheless, N-deglycosylated HF3 bound with higher affinity to collagens I and VI, although its proteolytic activity upon these collagens was not enhanced. This study demonstrates that features of carbohydrate moieties of haemorrhagic SVMPs may play a role in their interaction with substrates of the extracellular matrix, and the ability of SVMPs to degrade proteins in vitro does not correlate to their ability to cause haemorrhage, suggesting that novel, systemic approaches are necessary for understanding the mechanism of haemorrhage generation by SVMPs.
Oliveira
Ana K
AK
Laboratório Especial de Toxinologia Aplicada/CAT-CEPID, Instituto Butantan, São Paulo, Brazil.
Paes Leme
Adriana F
AF
Asega
Amanda F
AF
Camargo
Antonio C M
AC
Fox
Jay W
JW
Serrano
Solange M T
SM
D000818:Animals; D001798:Blood Proteins / Q000378:metabolism; D017837:Bothrops; D003435:Crotalid Venoms / Q000201:enzymology / Q000633:toxicity*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D006031:Glycosylation; D006470:Hemorrhage / Q000097:blood / Q000139:chemically induced*; D006801:Humans; D008666:Metalloendopeptidases / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D045726:Metalloproteases / Q000633:toxicity; D051379:Mice; D008970:Molecular Weight; D011485:Protein Binding; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D012871:Skin Diseases / Q000097:blood / Q000139:chemically induced*; D013329:Structure-Activity Relationship; D013997:Time Factors
D001798:Blood Proteins; D003435:Crotalid Venoms; D016326:Extracellular Matrix Proteins; D045726:Metalloproteases; C543527:HF3 protein, Bothrops jararaca; D008666:Metalloendopeptidases; C472493:bothropasin, Bothrops jararaca