Abstract
The type II cohesin domain and type II dockerin of bacterial cellulosome were cloned from Clostridium thermocellum and expressed with the fusion of tobacco mosaic virus coat protein (TMVcp) and enhanced green fluorescent protein (EGFP), respectively, in Escherichia coli. The TMVcp-cohesin fusion protein was assembled to the stable and rod-shaped nanostructure (TMVcp-Coh rod) under a particular buffer condition, where many active cohesin proteins are biologically and densely displayed around the 3-dimensional surface of TMVcp-Coh rod. Using EGFP-dockerin as a fluorescent reporter, we confirmed that the Ca2+-dependent binding and dissociation between native cohesin and dockerin were reproduced with the two recombinant fusion proteins, TMVcp-cohesin and EGFP-dockerin. The multi-cyclic binding-dissociation operation of TMVcp-Coh rod and EGFP-dockerin was successfully performed with maintaining the reversible cohesin-dockerin interaction in every cycle. EGFP that was fused to dockerin as a proof-of-concept here can be switched to other functional proteins/peptides that need to be used in multi-cyclic operation.
| Original language | English |
|---|---|
| Pages (from-to) | 101-106 |
| Number of pages | 6 |
| Journal | Journal of Biotechnology |
| Volume | 221 |
| DOIs | |
| Publication status | Published - 2016 Mar 10 |
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Keywords
- Cohesin
- Dockerin
- Multi-cyclic operation
- Reversible interaction
- Tobacco mosaic virus coat protein
ASJC Scopus subject areas
- Biotechnology
- Applied Microbiology and Biotechnology
Cite this
Reversible and multi-cyclic protein-protein interaction in bacterial cellulosome-mimic system using rod-shaped viral nanostructure. / Kim, Hyun Jin; Lee, Eun Jung; Park, Jin Seung; Sim, Sang Jun; Lee, Jeewon.
In: Journal of Biotechnology, Vol. 221, 10.03.2016, p. 101-106.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Reversible and multi-cyclic protein-protein interaction in bacterial cellulosome-mimic system using rod-shaped viral nanostructure
AU - Kim, Hyun Jin
AU - Lee, Eun Jung
AU - Park, Jin Seung
AU - Sim, Sang Jun
AU - Lee, Jeewon
PY - 2016/3/10
Y1 - 2016/3/10
N2 - The type II cohesin domain and type II dockerin of bacterial cellulosome were cloned from Clostridium thermocellum and expressed with the fusion of tobacco mosaic virus coat protein (TMVcp) and enhanced green fluorescent protein (EGFP), respectively, in Escherichia coli. The TMVcp-cohesin fusion protein was assembled to the stable and rod-shaped nanostructure (TMVcp-Coh rod) under a particular buffer condition, where many active cohesin proteins are biologically and densely displayed around the 3-dimensional surface of TMVcp-Coh rod. Using EGFP-dockerin as a fluorescent reporter, we confirmed that the Ca2+-dependent binding and dissociation between native cohesin and dockerin were reproduced with the two recombinant fusion proteins, TMVcp-cohesin and EGFP-dockerin. The multi-cyclic binding-dissociation operation of TMVcp-Coh rod and EGFP-dockerin was successfully performed with maintaining the reversible cohesin-dockerin interaction in every cycle. EGFP that was fused to dockerin as a proof-of-concept here can be switched to other functional proteins/peptides that need to be used in multi-cyclic operation.
AB - The type II cohesin domain and type II dockerin of bacterial cellulosome were cloned from Clostridium thermocellum and expressed with the fusion of tobacco mosaic virus coat protein (TMVcp) and enhanced green fluorescent protein (EGFP), respectively, in Escherichia coli. The TMVcp-cohesin fusion protein was assembled to the stable and rod-shaped nanostructure (TMVcp-Coh rod) under a particular buffer condition, where many active cohesin proteins are biologically and densely displayed around the 3-dimensional surface of TMVcp-Coh rod. Using EGFP-dockerin as a fluorescent reporter, we confirmed that the Ca2+-dependent binding and dissociation between native cohesin and dockerin were reproduced with the two recombinant fusion proteins, TMVcp-cohesin and EGFP-dockerin. The multi-cyclic binding-dissociation operation of TMVcp-Coh rod and EGFP-dockerin was successfully performed with maintaining the reversible cohesin-dockerin interaction in every cycle. EGFP that was fused to dockerin as a proof-of-concept here can be switched to other functional proteins/peptides that need to be used in multi-cyclic operation.
KW - Cohesin
KW - Dockerin
KW - Multi-cyclic operation
KW - Reversible interaction
KW - Tobacco mosaic virus coat protein
UR - http://www.scopus.com/inward/record.url?scp=84956695801&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84956695801&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2016.01.033
DO - 10.1016/j.jbiotec.2016.01.033
M3 - Article
C2 - 26820321
AN - SCOPUS:84956695801
VL - 221
SP - 101
EP - 106
JO - Journal of Biotechnology
JF - Journal of Biotechnology
SN - 0168-1656
ER -