Heterologous expression and affinity purification of eukaryotic membrane proteins in view of functional and structural studies: The example of the sarcoplasmic reticulum Ca(2+)-ATPase.

@article{Cardi2010HeterologousEA,
  title={Heterologous expression and affinity purification of eukaryotic membrane proteins in view of functional and structural studies: The example of the sarcoplasmic reticulum Ca(2+)-ATPase.},
  author={Delphine Cardi and C{\'e}dric Montigny and Bertrand Arnou and Marie Jidenko and Estelle Marchal and Marc le Maire and Christine Jaxel},
  journal={Methods in molecular biology},
  year={2010},
  volume={601},
  pages={247-67}
}
Heterologous SERCA1a Ca(2+)-ATPase (sarco-endoplasmic reticulum Ca(2+)-adenosine triphosphatase isoform 1a) from rabbit was expressed in yeast Saccharomyces cerevisiae as a fusion protein, with a biotin acceptor domain (BAD) linked to the SERCA C-terminus by a thrombin cleavage site. Thanks to the pYeDP60 vector, the recombinant protein was expressed under the control of a galactose-inducible promoter. Biotinylation of the protein occurred directly in yeast. Optimizing the number of galactose… CONTINUE READING