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Each protomer of a dimeric YidC functions as a single membrane insertase
Dirk Spann1, Eva Pross1, Yuanyuan Chen2
1Institute of Microbiology and Molecular Biology, University of Hohenheim, 70599, Stuttgart, Germany.
Scientific Reports
|January 14, 2018
Summary
The membrane insertase YidC, crucial for protein insertion, functions effectively as a dimer. This dimeric form allows YidC to act as two independent insertases, enhancing its membrane protein insertion capabilities.
Area of Science:
- Biochemistry
- Molecular Biology
- Membrane Biology
Background:
- YidC is an essential membrane insertase protein.
- YidC can exist as a monomer or dimer, or as part of the SecYEGDF-YajC-YidC holotranslocase complex.
- The functional role of dimeric YidC in protein insertion remains unclear.
Purpose of the Study:
- To investigate the functionality of dimeric YidC.
- To determine if two YidC protomers cooperate in inserting a single substrate.
- To explore the mechanism of YidC-mediated membrane protein insertion.
Main Methods:
- Construction of a fusion protein with two YidC copies linked together.
- Assessing the stability and functionality of the dimeric YidC fusion protein.
- Utilizing site-directed mutagenesis to inactivate individual YidC protomers.
- Monitoring substrate protein accumulation in the cytoplasm.
Main Results:
- The dimeric YidC fusion protein is stable and functional, supporting membrane insertion of various substrates.
- Mutations in one YidC protomer do not abolish insertase activity.
- Defects in both protomers lead to substrate protein accumulation in the cytoplasm.
- Dimeric YidC can bind two substrate proteins simultaneously.
Conclusions:
- Dimeric YidC functions as two independent insertases, rather than cooperating on a single substrate.
- The ability to bind two substrates simultaneously supports the independent function of each protomer.
- YidC's dimeric form enhances its capacity for membrane protein insertion.
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