Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization
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Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization. / Zedler, Julie A.Z.; Schirmacher, Alexandra M.; Russo, David A.; Hodgson, Lorna; Gundersen, Emil; Matthes, Annemarie; Frank, Stefanie; Verkade, Paul; Jensen, Poul Erik.
In: ACS Nano, Vol. 17, No. 24, 2023, p. 25279-25290.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization
AU - Zedler, Julie A.Z.
AU - Schirmacher, Alexandra M.
AU - Russo, David A.
AU - Hodgson, Lorna
AU - Gundersen, Emil
AU - Matthes, Annemarie
AU - Frank, Stefanie
AU - Verkade, Paul
AU - Jensen, Poul Erik
N1 - Publisher Copyright: © 2023 The Authors. Published by American Chemical Society.
PY - 2023
Y1 - 2023
N2 - Cyanobacteria offer great potential as alternative biotechnological hosts due to their photoautotrophic capacities. However, in comparison to established heterotrophic hosts, several key aspects, such as product titers, are still lagging behind. Nanobiotechnology is an emerging field with great potential to improve existing hosts, but so far, it has barely been explored in microbial photosynthetic systems. Here, we report the establishment of large proteinaceous nanofilaments in the unicellular model cyanobacterium Synechocystis sp. PCC 6803 and the fast-growing cyanobacterial strain Synechococcus elongatus UTEX 2973. Transmission electron microscopy and electron tomography demonstrated that expression of pduA*, encoding a modified bacterial microcompartment shell protein, led to the generation of bundles of longitudinally aligned nanofilaments in S. elongatus UTEX 2973 and shorter filamentous structures in Synechocystis sp. PCC 6803. Comparative proteomics showed that PduA* was at least 50 times more abundant than the second most abundant protein in the cell and that nanofilament assembly had only a minor impact on cellular metabolism. Finally, as a proof-of-concept for co-localization with the filaments, we targeted a fluorescent reporter protein, mCitrine, to PduA* by fusion with an encapsulation peptide that natively interacts with PduA. The establishment of nanofilaments in cyanobacterial cells is an important step toward cellular organization of heterologous pathways and the establishment of cyanobacteria as next-generation hosts.
AB - Cyanobacteria offer great potential as alternative biotechnological hosts due to their photoautotrophic capacities. However, in comparison to established heterotrophic hosts, several key aspects, such as product titers, are still lagging behind. Nanobiotechnology is an emerging field with great potential to improve existing hosts, but so far, it has barely been explored in microbial photosynthetic systems. Here, we report the establishment of large proteinaceous nanofilaments in the unicellular model cyanobacterium Synechocystis sp. PCC 6803 and the fast-growing cyanobacterial strain Synechococcus elongatus UTEX 2973. Transmission electron microscopy and electron tomography demonstrated that expression of pduA*, encoding a modified bacterial microcompartment shell protein, led to the generation of bundles of longitudinally aligned nanofilaments in S. elongatus UTEX 2973 and shorter filamentous structures in Synechocystis sp. PCC 6803. Comparative proteomics showed that PduA* was at least 50 times more abundant than the second most abundant protein in the cell and that nanofilament assembly had only a minor impact on cellular metabolism. Finally, as a proof-of-concept for co-localization with the filaments, we targeted a fluorescent reporter protein, mCitrine, to PduA* by fusion with an encapsulation peptide that natively interacts with PduA. The establishment of nanofilaments in cyanobacterial cells is an important step toward cellular organization of heterologous pathways and the establishment of cyanobacteria as next-generation hosts.
KW - cyanobacteria
KW - encapsulation peptide
KW - nanofilament
KW - PduA
KW - protein scaffold
KW - Synechococcus elongatus UTEX 2973
KW - Synechocystis sp. PCC 6803
U2 - 10.1021/acsnano.3c08600
DO - 10.1021/acsnano.3c08600
M3 - Journal article
C2 - 38065569
AN - SCOPUS:85180116039
VL - 17
SP - 25279
EP - 25290
JO - A C S Nano
JF - A C S Nano
SN - 1936-0851
IS - 24
ER -
ID: 380206787