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Master's Dissertation
DOI
10.11606/D.9.2011.tde-20052013-165307
Document
Author
Full name
Edson Mendes de Oliveira
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Campa, Ana (President)
Cintra, Dennys Esper Corrêa
Fock, Ricardo Ambrosio
Title in Portuguese
Amilóide sérica A (SAA): produção da proteína recombinante humana SAA1 e SAA4 e sua expressão nativa em células do tecido adiposo submetidas à hipóxia
Keywords in Portuguese
Adipócito
Hipóxia
Pré-adipócito 3T3-L1
Proteínas recombinantes
SAA
Abstract in Portuguese
Visando novos estudos com a proteína amilóide sérica A (SAA), propusemos a produção de seu recombinante humano em Escherichia coli, mais especificamente, a isoforma encontrada na fase aguda (A-SAA1) e da constitutivamente expressa (C-SAA4). Realizamos a expressão, identificação e purificação das proteínas recombinantes. Concomitantemente, também avaliamos o efeito da hipóxia na expressão e produção da proteína SAA nativa em linhagens de pré-adipócitos murinos 3T3-L1, não diferenciados e diferenciados e adipócitos humanos. Aparentemente quanto maior o grau de diferenciação celular, maior a expressão e produção da proteína. Para os adipócitos humanos, o perfil de expressão de mRNA da SAA mostra que SAA1>SAA2>SAA4 nas relações 500:150:1. Na hipóxia, há um aumento na expressão de SAA, entretanto não associamos esta expressão a um aumento da concentração da proteína. A importância do reconhecimento de que SAA pode ser umas das proteínas induzidas pela hipóxia em adipócitos é discutida em relação ao seu papel pró-inflamatório.
Title in English
Serum amyloid A (SAA): production of recombinant human protein SAA1 and SAA4 and its native expression on adipose tissue cells submitted to hypoxia
Keywords in English
3T3-L1 preadipocytes
Adipocyte
Hypoxia
Recombinant protein
SAA
Abstract in English
In order to provide more complex studies with the protein serum amyloid A (SAA), this study proposed the production of its human recombinant in bacteria (Escherichia coli), more specifically, the synthesis of the main isoform found in the acute phase (A-SAA1) and the constitutively expressed (C-SAA4). The expression, identification and purification of the recombinants proteins was performed. Concurrently, we also did a study evaluating the effect of hypoxia in the expression and protein production of native SAA in undifferentiated and differentiated murine preadipocytes 3T3-L1 and humans adipocytes. Apparently, the protein expression and production increases with the cell differentiation degree. For human adipocytes, we demonstrated the mRNA expression profile of SAA, in which SAA1 > SAA2 > SAA4 (500:150:1). In hypoxia, there is an increased expression of SAA, but we could not link it to an increase of protein concentration. The importance of recognizing that SAA may be one of the proteins induced by hypoxia in adipocytes is discussed in relation to the proinflammatory role of this protein.
 
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Publishing Date
2013-07-03
 
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