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Published online before print June 6, 2008
Protein Science, DOI: 10.1110/ps.035576.108
 Copyright © 2008 The Protein Society
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Enhanced protein expression in mammalian cells using engineered SUMO fusions: Secreted Phospholipase A2

Raymond J. Peroutka1, Nabil Elshourbagy1, Tara L. Piech1, and Tauseef R. Butt2,3

1 LifeSensors, Inc.;
2 LifeSensors.com

(RECEIVED April 3, 2008; ACCEPTED June 4, 2008)

SUMOylation, the covalent attachment of SUMO (small ubiquitin-like modifier), is a eukaryotic posttranslational event that has been demonstrated to play a critical role in several biological processes. When used as an N-terminal tag or fusion partner, SUMO has been shown to enhance functional protein production significantly by improving folding, solubility and stability. We have engineered several SUMOs and, through their fusion, developed a system for enhancing the expression and secretion of complex proteins. To demonstrate the fidelity of this fusion technology, secreted phospholipase A2 proteins (sPLA2) were produced using HEK-293 and CHO-K1 mammalian cells. Four mouse sPLA2 homologues were expressed and secreted in mammalian cell cultures using the SUMO or SUMO-derived, N-terminal fusion partners. Mean and median increases of 43 fold and 18 fold, respectively, were obtained using novel SUMO mutants that are resistant to digestion by endogenous desumoylases.

Keywords: Protein Structure/Folding; Stability and mutagenesis; Enzymes of arachidonate metabolism; Purification methods, general; cDNA, Mutagenesis, Cloning; Mutagenesis (site-directed and general); Expression systems; SUMO

3 E-mail: butt{at}lifesensors.com


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