TY  - JOUR
A1  - Bourdeaux, Florian Alexander
A1  - Kopp, Yannick
A1  - Lautenschläger, Julia
A1  - Gößner, Ines
A1  - Besir, Hüseyin
A1  - Vabulas, R. Martin
A1  - Grininger, Martin
T1  - Dodecin as carrier protein for immunizations and bioengineering applications
T2  - Scientific reports
N2  - In bioengineering, scaffold proteins have been increasingly used to recruit molecules to parts of a cell, or to enhance the efficacy of biosynthetic or signalling pathways. For example, scaffolds can be used to make weak or non-immunogenic small molecules immunogenic by attaching them to the scaffold, in this role called carrier. Here, we present the dodecin from Mycobacterium tuberculosis (mtDod) as a new scaffold protein. MtDod is a homododecameric complex of spherical shape, high stability and robust assembly, which allows the attachment of cargo at its surface. We show that mtDod, either directly loaded with cargo or equipped with domains for non-covalent and covalent loading of cargo, can be produced recombinantly in high quantity and quality in Escherichia coli. Fusions of mtDod with proteins of up to four times the size of mtDod, e.g. with monomeric superfolder green fluorescent protein creating a 437 kDa large dodecamer, were successfully purified, showing mtDod’s ability to function as recruitment hub. Further, mtDod equipped with SYNZIP and SpyCatcher domains for post-translational recruitment of cargo was prepared of which the mtDod/SpyCatcher system proved to be particularly useful. In a case study, we finally show that mtDod-peptide fusions allow producing antibodies against human heat shock proteins and the C-terminus of heat shock cognate 70 interacting protein (CHIP).
KW  - Biotechnology
KW  - Proteins
Y1  - 2020
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/80310
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-803102
SN  - 2045-2322
N1  - This work was supported by intramural funds of the Goethe University Frankfurt to M.G., and was associated to the LOEWE program (Landes-Offensive zur Entwicklung wissenschaftlich-ökonomischer Exzellenz) of the state of Hesse conducted within the framework of the MegaSyn Research Cluster. Open access funding provided by Projekt DEAL.
VL  - 10
IS  - art. 13297
SP  - 1
EP  - 18
PB  - Macmillan Publishers Limited, part of Springer Nature
CY  - [London]
ER  -