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New Biological Frontiers Illuminated by Molecular Sensors and Actuators
Monday Speaker Abstracts
Optomechanical Actuators for Controlling Mechanotransduction in Living Cells
Khalid Salaita
1,2
.
1
Emory University, Atlanta, USA,
2
Georgia Institute of Technology & Emory University,
Atlanta, GA, USA.
See abstract: Pos-37 Board 37
Synthetically Rerouting Phagocytosis by Rapidly Turning Inert Cells into “Eat You” Mode
Toru Komatsu
1,4
, Hiroki Onuma
1
, Tetsuo Nagano
2
, Yasuteru Urano
1,3
.Takanari Inoue
5
.
1
The University of Tokyo, Tokyo, Japan,
3
The University of Tokyo, Tokyo, Japan,
4
JST
PRESTO, Tokyo, Japan,
5
Johns Hopkins University, Baltimore, MD, USA.
2
The University of
Tokyo, Tokyo, Japan,
See Abstract: Pos-19 Board 19
Organization of Intracellular Reactions with Heterologous Protein Scaffold
Hsiao-Chun Huang
.
National Taiwan University, Taipei, Taiwan.
Prokaryotes lack membranous organelles to compartmentalize biochemical reactions. To
optimize the efficiency of engineered metabolic pathways in E.coli, artificial organelles based on
porous protein shells and synthetic RNA/protein scaffold have been proposed to bring metabolic
enzymes to close proximity and thus speed up reactions. In this study, we aim to develop, in
E.coli, a unique protein scaffold to spatially assemble pathways of interest, based on
heterologous expression of Caulobacter crescentus proteins, PopZ and SpmX. We have cloned
and standardized both PopZ and SpmX genes from Caulobacter crescentus, and created
fluorescent fusions to validate their localization and capability as molecular scaffold in E.coli.
We scanned different expression levels of PopZ and established stable formation of PopZ
complex in E.coli. We found that SpmX remained diffused throughout when it was singly
expressed in E.coli. When SpmX was co-expressed with PopZ, it then co-localized with PopZ
foci, suggesting that there is direct interaction between PopZ and SpmX. We then tested if SpmX
can serve as the adaptor for PopZ to bring different proteins of interest to close proximity. As a
proof of concept, we performed bimolecular fluorescence complementation. No fluorescence
was observed when these two split fluorescent fusion proteins were co-expressed. Fluorescent
foci was detected when PopZ is present, strongly suggesting that PopZ complex can serve as a
molecular scaffold to spatially assemble pathways via SpmX∆C adaptors. With this scaffold
device, we plan to assemble and optimize the efficiency of a foreign metabolic pathway in E.coli.
We anticipate this system to be user-friendly in wide range of microbial metabolic engineering
applications.