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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2015.tde-10122015-074655
Document
Author
Full name
Daniela Tiemi Myamoto
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2015
Supervisor
Committee
Tambourgi, Denise Vilarinho (President)
Aniz, Patricia Antonia Estima Abreu de
Azevedo, Inácio de Loiola Meirelles Junqueira de
Barbosa, Ângela Silva
Isaac, Lourdes
Title in Portuguese
Caracterização e análise filogenética dos genes que codificam para os componentes C3 e fator B do sistema complemento das glândulas de veneno de aranhas Loxosceles.
Keywords in Portuguese
Loxosceles laeta
C3
Clonagem
FB
Filogenia
Sistema complemento
Abstract in Portuguese
O sistema complemento parece ter surgido com o aparecimento de C3 e fator B (FB), os únicos componentes encontrados nos organismos mais primitivos, o que sugere que a via alternativa seja a mais antiga. Fragmentos de cDNA codificantes para FB (Lox-FB) e C3 (Lox-C3) foram sintetizados a partir do RNA total isolado da glândula de veneno de Loxosceles laeta e amplificados por técnicas de RACE-PCR. Lox-FB apresenta uma organização de domínios clássica, composta por domínios CCP, vWFA e de serino protease. Os aminoácidos envolvidos na ligação ao C3b são conservados, no entanto, a tríade catalítica clássica não foi encontrada. Lox-C3 apresenta uma configuração de domínios similar a do C3 humano, contendo dois sítios putativos de processamento: um entre as cadeias α e γ, e outro entre as cadeias α e β, indicando que Lox-C3 seja composto por três cadeias. As análises filogenéticas indicaram que Lox-C3 e Lox-FB são mais próximos evolutivamente aos componentes equivalentes da aranha Hasarius adansoni, com valores de identidade de 53% e 43%, respectivamente.
Title in English
Characterization and phylogenetic analysis of genes coding for the components C3 and factor B of Complement System from Loxosceles spiders venom glands.
Keywords in English
Loxosceles laeta
C3
Cloning
Complement system
FB
Phylogeny
Abstract in English
The complement system seems to have arisen with the appearance of C3 and factor B (FB), the only components found in the most primitive organisms, suggesting that the alternative pathway is the oldest. cDNA fragments coding for the complement factor B (Lox-FB) and C3 (Lox-C3) were synthesized from total RNA Loxosceles laeta venom gland and amplified using RACE-PCR techniques. Lox-FB has a classical domain organization in mosaic, composed by CCPs, vWFA and serine protease domains. The amino acids involved in binding to C3b are conserved, however, the classical calatytic triad was not found. Lox-C3 presents a similar configuration of domains to C3 human and has two putative processing sites: the first one is located between α and γ chains and other between α and β chains, indicating that Lox-C3 is composed by three chains. The phylogenetic analyses indicated that Lox-C3 and Lox-FB are evolutionary closer to the equivalent components of Hasarius adansoni, with identity values of 53% and 43%, respectively.
 
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Publishing Date
2015-12-10
 
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