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Published on: February 17, 2014
Predicted bacteriorhodopsin from Exiguobacterium sibiricum is a functional proton pump.
L E Petrovskaya1, E P Lukashev, V V Chupin
1Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russian Federation. lpetr65@yahoo.com
Researchers expressed Exiguobacterium sibiricum bacteriorhodopsin (ESR) from ancient permafrost, finding it binds retinal and transports protons. This ancient protein functions in a proton pump, offering insights into microbial adaptation.
Area of Science:
- Microbiology
- Biochemistry
- Molecular Biology
Background:
- Bacteriorhodopsins are microbial proton pumps.
- Ancient DNA offers insights into evolutionary biology.
- Permafrost preserves ancient genetic material.
Purpose of the Study:
- To characterize the bacteriorhodopsin from Exiguobacterium sibiricum (ESR).
- To investigate the functional properties of ESR.
- To explore the potential of ancient microbial proteins.
Main Methods:
- Gene amplification from ancient Siberian permafrost.
- Protein expression in Escherichia coli.
- Spectroscopic analysis (absorbance).
- Functional assays using proteoliposomes (proton transport).
Main Results:
- ESR gene successfully amplified and protein expressed.
- ESR bound all-trans-retinal, showing a 534nm absorbance maximum.
- ESR exhibited a photocycle with rapid M intermediate formation and a stable O intermediate.
- ESR-containing proteoliposomes demonstrated outward proton transport, acidifying the medium.
Conclusions:
- Exiguobacterium sibiricum bacteriorhodopsin is a functional proton pump.
- ESR's photocycle characteristics suggest efficient proton translocation.
- This ancient protein offers a model for studying microbial energy transduction and adaptation.
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