Cloning, expression and characterization of molecules coming from the salivary complex of leeches Haementeria gender.

• Grant number: 16/09137-7
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: July 01, 2016
• End date: December 31, 2019
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Ana Marisa Chudzinski-Tavassi
• Grantee: Adriane Michele Xavier Prado Amorim
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil

Abstract

Bloodsucking animals developed a wide range of substances that are injected into the host through saliva and inhibit the hemostatic reactions at the site where they feed. With this, various components have been described with activities in different haemostatic processes (coagulation, fibrinolysis and platelet aggregation).Leeches Haementeria vizottoi and Haementeria depressa have been studied by our group and by classical protein biochemistry, from complex salivary these leeches, proteins with interesting roles in hemostasis were characterized, among them stand out: the Vizottin a factor inhibitor Xa (Oliveira et al, 2012) salivary complexes H. vizottoi; Lefaxin an inhibitor of the blood coagulation pathway FXa (Faria et al., 1999); the Hementerin initially characterized as fibrin (geno) lytic (Chudzinski-Tavassi et al., 1998) and subsequently also described as an inhibitor of platelet aggregation by nitridérgica means (Chudzinski-Tavassi et al., 2003), the latter being from the salivary complex H. depressa.Recently, it performed the transcriptomic study of salivary complex of leeches H.vizottoi, the profile of transcripts was determined where 1747 sequences were obtained (Amorim et al., 2015). Among the transcripts involved in the feeding process, the Isotig234, showed 71% similarity with Antistasin (Nutt et al., 1 988) and 75% similarity with the ghilanten (Blankenship et al., 1 990) and Isotig1411 presented 34% identity with the platelet aggregation inhibitor (LAPP) (Connoly et al., 1992).Through a transcriptomic and proteomic analysis of salivary complex leech H. depressa, the profile of transcripts and proteins produced by the tissue was determined (Faria et al., 2005; Ricci-Silva et al., 2005), transcribed from the L01C01 clone showed 92% similarity with the tridegin a FXIIIa inhibitor from leeches Haementeria ghilianii (Seale et al., 1997).A large number of synthetic and recombinant molecules has been obtained based on natural inhibitors. The saliva of bloodsucking animals such as leeches, has been focused to the discovery of new molecules that may have a role in anticoagulation.The present work proposes, the expression and characterization of recombinant inhibitors antistasin like and platelet aggregation Brazilian leech H. vizottoi and an FXIIIa inhibitor leech H. depressa. These types of inhibitors are molecules that are important in the global therapeutic scenario due to its high pharmaceutical potential hemostatic disorders, which comes praise our study.