Produção Nacional
Revisado por pares
Lopap, a prothrombin activator from Lonomia obliqua belonging to the lipocalin family: recombinant production, biochemical characterization and structure–function insights
2006; Portland Press; Volume: 398; Issue: 2 Linguagem: Inglês
10.1042/bj20060325
ISSN1470-8728
AutoresCleyson Valença Reis, Sonia Aparecida de Andrade, Oscar Henrique Pereira Ramos, Celso Raul Romero Ramos, Paulo Lee Ho, Isabel de Fátima Correia Batista, Ana Marisa Chudzinski‐Tavassi,
Tópico(s)Venomous Animal Envenomation and Studies
ResumoUsing a cDNA library made from Lonomia obliqua caterpillar bristles, we identified a transcript with a 603 bp open reading frame. The deduced protein corresponds to Lopap, a prothrombin activator previously isolated by our group from the bristles of this species. The mature protein is composed by 185 amino acids and shares similarity with members of the lipocalin family. The cDNA encoding the mature form was amplified by PCR, subcloned into pAE vector and used to transform Escherichia coli BL21(DE3) cells. As for the native Lopap, the recombinant fusion protein shows enzymatic activity, promotes prothrombin hydrolysis, generates fragments similar to prethrombin-2 and fragment 1.2 as intermediates, and generates thrombin as the final product. In addition, structural bioinformatics studies indicated several interesting molecular features, including the residues that could be responsible for Lopap's serine protease-like activity and the role of calcium binding in this context. Such catalytic activity has never been found in other members of the lipocalin family. This is the first report describing the recombinant production and biochemical characterization of a Lonomia obliqua lipocalin, as well as the structural features that could be responsible for its serine protease-like catalytic activity.
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