Navigation Links
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.

Under oxidative conditions intramolecular disulfide bonds can form, shifting the free energy minimum from the α-helix, to a clamped-coil state (similar to a helix-coil transition). The coiled state allows the two fluorescent proteins to approach closer than in the extended helix state, where they can more efficiently exchange excitation energy (i.e., a high FRET state). The extent of energy transfer is easily quantified from the increased emission of the acceptor.

This is the first step towards development of a FRET-based biosensor for visualizing redox potentials and oxidative stress in live cells and tissues via optical microscopy.

We employed a sensitive technique for measuring FRET to screen our linkers. This methodology greatly expedited the quantitative analysis and development of the linkers and will be very useful for the development of other FRET-based sensors, said Bryan Q. Spring, a doctoral student and co-author of the publication. Given the importance of the intracellular redox state in determining a cells fate, and the increasing evidence that perturbations in the redox state are associated with cancer and various inflammatory disorders as well as aging, FRET-based redox sensors offer significant promise for understanding molecular mechanisms underlying human health and disease.

Dr. Steven R. Goodman, Editor-in-Chief of Experimental Biology and Medicine, said Altered redox status is a hallmark of many diseases ranging from neurological disorders, such as Alzheimers Disease, to hematologic disorders such as Sickle Cell Disease. The development of a FRET-based biosensor to measure oxidative stress in living cells would be of enormous benefit to biomedical researchers working in many diverse fields. This is precisely the type of interdisciplinary effort that the new Experimental Biology and Medicine hopes to provide to the international scientific community.


'/>"/>

Contact: Dr. Vladimir L. Kolossov
viadimer@uiuc.edu
217-244-3163
Society for Experimental Biology and Medicine
Source:Eurekalert  

Related medicine news :

1. MSU engineering team designs innovative medical device
2. Specialty Blades, Inc. Expands Management Team With New VP Engineering
3. McGraw-Hill Professional Enhances Library Access to Medical and Engineering Resources
4. Riverside Partners Teams with Management to Acquire GEMCITY Engineering & Manufacturing
5. Bayer Foundation Grant Creates Graduate Engineering Fellowship at University of Houston
6. American Oriental Bioengineering Announces Participation in November Healthcare Conferences
7. Springer to publish Sports Engineering
8. American Oriental Bioengineering to Present at the ICR XChange Asia Institutional Investor Conference
9. Antibody Engineering Company f-star Appoints Three Pioneers in Antibody Engineering and Development to its Newly Established Scientific Advisory Board
10. CDI-Life Sciences Selected by CJ CheilJedang Corp. to Provide Engineering Design for Major Pharmaceutical Complex in Korea
11. MedImmune Expands Scientific and Engineering Leadership
Post Your Comments:
*Name:
*Comment:
*Email:
Related Image:
Engineering chimeric polypeptides to illuminate cellular redox states
(Date:6/25/2016)... ... June 25, 2016 , ... Experts from ... at AcademyHealth’s Annual Research Meeting June 26-28, 2016, at the Hynes Convention Center ... care topics including advance care planning, healthcare costs and patient and family engagement. ...
(Date:6/25/2016)... TX (PRWEB) , ... June 25, 2016 , ... ... the United States, named Dr. Sesan Ogunleye, as the Medical Director of its new ... the facility Medical Director of our new Mesquite location,” said Dr. James M. Muzzarelli, ...
(Date:6/25/2016)... ... June 25, 2016 , ... Conventional wisdom preaches the ... In terms of the latter, setting the bar too high can result in disappointment, ... just slow progress toward their goal. , Research from PsychTests.com reveals ...
(Date:6/24/2016)... ... ... crisis. Her son James, eight, was out of control. Prone to extreme mood shifts and ... him, he couldn’t control his emotions,” remembers Marcy. “If there was a knife on ... say he was going to kill them. If we were driving on the freeway, ...
(Date:6/24/2016)... ... 24, 2016 , ... Comfort Keepers® of San Diego, CA is excited to ... Recovery® program to drive cancer patients to and from their cancer treatments. Comfort ... quality of life and ongoing independence. Getting to and from medical treatments is ...
Breaking Medicine News(10 mins):
(Date:6/24/2016)... SOUTH SAN FRANCISCO, Calif. , June 24, ... GBT ), a biopharmaceutical company developing novel ... with significant unmet needs, today announced the closing ... 6,400,000 shares of common stock, at the public ... the shares in the offering were offered by ...
(Date:6/24/2016)... 24, 2016 ... the addition of the " Global Markets for ... This report focuses on ... updated review, including its applications in various applications. The ... which includes three main industries: pharmaceutical and biotechnology, food ...
(Date:6/24/2016)... ALEXANDRIA, Va. , June 24, 2016 ... a set of recommendations that would allow ... information (HCEI) with entities that make formulary and coverage ... determine the "value" of new medicines. The ... that does not appear on the drug label, a ...
Breaking Medicine Technology: