Universität zu Köln

Reuten, Raphael ORCID: 0000-0002-9434-4108, Nikodemus, Denise ORCID: 0000-0003-4492-817X, Oliveira, Maria B., Patel, Trushar R. ORCID: 0000-0003-0627-2923, Brachvogel, Bent, Breloy, Isabelle, Stetefeld, Joerg ORCID: 0000-0003-1478-3248 and Koch, Manuel (2016). Maltose-Binding Protein (MBP), a Secretion-Enhancing Tag for Mammalian Protein Expression Systems. PLoS One, 11 (3). SAN FRANCISCO: PUBLIC LIBRARY SCIENCE. ISSN 1932-6203

Full text not available from this repository.

Abstract

Recombinant proteins are commonly expressed in eukaryotic expression systems to ensure the formation of disulfide bridges and proper glycosylation. Although many proteins can be expressed easily, some proteins, sub-domains, and mutant protein versions can cause problems. Here, we investigated expression levels of recombinant extracellular, intracellular as well as transmembrane proteins tethered to different polypeptides in mammalian cell lines. Strikingly, fusion of proteins to the prokaryotic maltose-binding protein (MBP) generally enhanced protein production. MBP fusion proteins consistently exhibited the most robust increase in protein production in comparison to commonly used tags, e.g., the Fc, Glutathione S-transferase (GST), SlyD, and serum albumin (ser alb) tag. Moreover, proteins tethered to MBP revealed reduced numbers of dying cells upon transient transfection. In contrast to the Fc tag, MBP is a stable monomer and does not promote protein aggregation. Therefore, the MBP tag does not induce artificial dimerization of tethered proteins and provides a beneficial fusion tag for binding as well as cell adhesion studies. Using MBP we were able to secret a disease causing laminin beta 2 mutant protein (congenital nephrotic syndrome), which is normally retained in the endoplasmic reticulum. In summary, this study establishes MBP as a versatile expression tag for protein production in eukaryotic expression systems.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Reuten, Raphael UNSPECIFIED orcid.org/0000-0002-9434-4108 UNSPECIFIED Nikodemus, Denise UNSPECIFIED orcid.org/0000-0003-4492-817X UNSPECIFIED Oliveira, Maria B. UNSPECIFIED UNSPECIFIED UNSPECIFIED Patel, Trushar R. UNSPECIFIED orcid.org/0000-0003-0627-2923 UNSPECIFIED Brachvogel, Bent UNSPECIFIED UNSPECIFIED UNSPECIFIED Breloy, Isabelle UNSPECIFIED UNSPECIFIED UNSPECIFIED Stetefeld, Joerg UNSPECIFIED orcid.org/0000-0003-1478-3248 UNSPECIFIED Koch, Manuel UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-280973
DOI:	10.1371/journal.pone.0152386
Journal or Publication Title:	PLoS One
Volume:	11
Number:	3
Date:	2016
Publisher:	PUBLIC LIBRARY SCIENCE
Place of Publication:	SAN FRANCISCO
ISSN:	1932-6203
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language HIGH-LEVEL EXPRESSION; ESCHERICHIA-COLI; FUSION PROTEINS; DOMAIN; CELLS; ACID Multiple languages Multidisciplinary Sciences Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/28097