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Engineering chimeric polypeptides to illuminate cellular redox states
Date:1/23/2008

UIUC interdisciplinary team reports the design of chimeric polypeptides leading to development of noninvasive biosensors for potential application in biomedical research.

Reduction/oxidation (redox) systems research is reaching a stage where domains that traditionally belonged to the physical sciences, chemistry, and molecular biology are coming together to offer new synergistic opportunities for understanding and manipulating basic cellular processes that underlie complex biomedical problems (e.g., tumorigenesis). Parallel with this advance is the emerging recognition that the intracellular redox environment exerts a profound influence on the normal cellular processes of DNA synthesis, enzyme activation, selective gene expression, cell cycle progression, proliferation, differentiation, and apoptosis. However, this is a difficult area of study and molecular mechanisms mediating redox sensitivity are poorly defined.

An interdisciplinary research team from the University of Illinois Institute for Genomic Biology (IGB) report in the February issue of the journal Experimental Biology and Medicine the engineering of novel peptide sequences that are sensitive to redox conditions inside cells.

Attachment of linkers between a special pair of green fluorescent proteins (GFP) shows great promise for developing genetically encoded redox sensitive biosensors, said Vladimir L. Kolossov, corresponding author. To detect oxidation and reduction, the biosensor uses a powerful optical technique called Frster resonance energy transfer (FRET).

The absence of polypeptide linkers able to sense the redox state by undergoing a conformational change was the major obstacle to a FRET-based redox sensor. The researchers designed the linker sequence such that in its reduced state the linker is an α-helix. Thiol groups, strategically placed throughout the linker, sense the redox potential of the environment and form disulfide bonds upon oxidation
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Contact: Dr. Vladimir L. Kolossov
viadimer@uiuc.edu
217-244-3163
Society for Experimental Biology and Medicine
Source:Eurekalert  

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