Abstract
Aggresomes are protein aggregates found in mammalian cells when the intracellular protein degradation machinery is over-titered. Despite that they abound in cells producing recombinant proteins of biomedical and biotechnological interest, the physiological roles of these protein clusters and the functional status of the embedded proteins remain basically unexplored. In this work, we have determined for the first time that, like in bacterial inclusion bodies, deposition of recombinant proteins into aggresomes does not imply functional inactivation. As a model, human α-galactosidase A (GLA) has been expressed in mammalian cells as enzymatically active, mechanically stable aggresomes showing higher thermal stability than the soluble GLA version. Since aggresomes are easily produced and purified, we propose these particles as novel functional biomaterials with potential as carrier-free, self-immobilized catalyzers in biotechnology and biomedicine.
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Acknowledgments
MALDI-TOF analyses were carried out in the Proteomics and Structural Biology Facility (SepBioEs) from UAB, member of ProteoRed network. We thank the Protein Expression Core Facility, Institute for Research in Biomedicine (IRB, Barcelona) for cloning the GLA gene into the pOPIN vector. We appreciate the technical support from the UAB Scientific and Technical Services SCAC (Servei de Cultius Cel · lulars, Producció d'Anticossos i Citometria). Protein production has been partially performed by the ICTS “NANBIOSIS,” more specifically by the Protein Production Platform of CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER-BBN)/IBB, at the UAB (http://www.ciber-bbn.es/en/programas/89-plataforma-de-produccion-de-proteinas-ppp). Authors appreciate the financial support through the grant EUI2008-03610 (MICINN) linked to the ERANET-IB 08–007 project, from Agència de Gestió d’Ajuts Universitaris i de Recerca (2014SGR-132), and for the grant to the project “Development of nanomedicinas for enzymatic replacement therapy in Fabry disease,” granted by the Fundació Marató TV3. We also acknowledge the support of the CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. AV was distinguished with an ICREA ACADEMIA award (Catalonia, Spain).
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Rodríguez-Carmona, E., Mendoza, R., Ruiz-Cánovas, E. et al. A novel bio-functional material based on mammalian cell aggresomes. Appl Microbiol Biotechnol 99, 7079–7088 (2015). https://doi.org/10.1007/s00253-015-6684-0
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DOI: https://doi.org/10.1007/s00253-015-6684-0