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Doctoral Thesis
DOI
10.11606/T.46.1974.tde-23072012-155915
Document
Author
Full name
Maria Julia Manso Alves
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 1974
Supervisor
Title in Portuguese
Isolamento e purificação de glicoproteínas de Trypanossoma cruzi (epimastigota)
Keywords in Portuguese
Bioquímica
Epimastigota
Galactofuranose
Glicoproteínas
Trypanosoma cruzi
Abstract in Portuguese
Forma epimastigotas de T. cruzi são aglutinadas especificamente por baixas concentrações de concanavalina A. A aglutinação é linear com o tempo até aproximadamente10 minutos, para um número de células maior que 1 x 108 células/ml. Nessas condições a aglutinação é dependente da concentração de con A. As formas tripomastigotas sanguícolas e matecíclicas não são aglutináveis por con A. O complexo glicoproteico isolado de formas epimastigotas de T. cruzi por tratramentofenólico de extrato celular, apresenta na sua composição ácida siálico, glicosamina, galactose, glicose e manose, além de xilose em algumas frações. Os aminoácidos constituintes são principalmente lisina, ácido aspártico (e/ou asparagina), alanina, treonina, ácido glutâmico (e/ou glutamina). serina, prolina e glicina. Esse complexo pode ser separado em três componentes em colunas de DEAE-celulose. Dois desses componentes, selecionados para estudo, inibem a reação de aglutinação por con A das formas epimastigotas, assim como a fração não cromatografada. Essas frações isoladas fornecem quatro componentes glicoproteicos por eletroforese em gel de poliacrilamida na presença de SDS.
Title in English
Isolation and purification of glycoproteins from Trypanosoma cruzi (epimastigotes)
Keywords in English
Biochemistry
Epimastigote
Galactofuranose
Glycoproteins
Trypanosoma cruzi
Abstract in English
Epimastigote forms of T. cruzi are agglutinated by low con A concentrations. The agglutinabillity is linear up to 10 minutes. Under these conditions the agglutination is dependent on the con A concentration providing the cell density is 108/ml or higher. The blood forms and culture trypomastigotes are not agglutinated by con A. A glycoprotein complex was isolated from epimastigote forms of T. cruzi by aqueous phenol extraction. This complex is composed by sialic acid, glucosamine, galactose, glucose, mannose and xylose. The latter appears only in some fractions of the complex. The amino acids found are lysine, aspartic acid (and/or asparagine), alanine, threonine, glutamic acid (and/or glutamine), serine, proline and glycine. The glycoproteins render three components in DEAE-cellulose columns (peaks 1, 2 and 3). The peaks 2 and 3 are able to inhibit the agglutination of epimastigotes by con A. These fractions are separated in four glycoprotein components by polyacrylamide gel electrophoresis in the presence of SDS.
 
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Publishing Date
2012-07-23
 
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