Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line / 亚洲男科学杂志(英文版)
Asian Journal of Andrology
;
(6): 3-13, 2004.
Artigo
em Inglês
| WPRIM
| ID: wpr-300872
ABSTRACT
<p><b>AIM</b>To produce biologically active recombinant human (rh) ZP proteins in a human cell for use in sperm function tests.</p><p><b>METHODS</b>The human embryonic kidney cell line 293T was employed to produce rhZP1, rhZP2 and rhZP3 proteins individually and together by co-expression. Presence of these proteins in the culture medium and cell lysate was assessed by Western blotting analysis. The effect of the recombinant proteins on the human AR was assessed.</p><p><b>RESULTS</b>RhZP2 and rhZP3 were secreted into the culture medium, whereas rhZP1 was found only in the cell lysate. Interestingly, when all zona pellucida proteins were co-expressed in the same cells, rhZP1 was also secreted into the culture medium. However, despite the presence of all three ZP proteins in sufficient concentration and evidence of heavy glycosylation on gel electrophoresis, biological activity to induce the AR was not observed.</p><p><b>CONCLUSION</b>RhZP1, rhZP2 and rhZP3 were successfully expressed in the human embryonic kidney cell line 293T. It appears that an interaction amongst these proteins may be required for release of rhZP1 from the cell. Although this approach is not satisfactory for producing active human ZP proteins, it makes a significant contribution to the understanding of the structural and functional characteristics of the ZP proteins.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Farmacologia
/
Glicosilação
/
Proteínas Recombinantes
/
Glicoproteínas de Membrana
/
Proteínas do Ovo
/
Expressão Gênica
/
Linhagem Celular
/
Western Blotting
/
Imunofluorescência
/
Receptores de Superfície Celular
Limite:
Feminino
/
Humanos
/
Masculino
Idioma:
Inglês
Revista:
Asian Journal of Andrology
Ano de publicação:
2004
Tipo de documento:
Artigo
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