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:9/20/2017)... Raton (PRWEB) , ... September 20, 2017 , ... In ... Daily Brain Booster has already been receiving positive feedback from customers trying the product ... months. , Daily Brain Booster was developed by neurosurgeon Shawn Moore, MD, for ...
(Date:9/20/2017)... , ... September 20, 2017 , ... ... of older Americans at risk of price gouging for their prescription drugs, according ... (TSCL) . “Because Medicare isn’t negotiating on our behalf, there’s no consistency in ...
(Date:9/20/2017)... ... September 20, 2017 , ... ... “RECYCLED Ezekiel's Plan for Freedom from ADDICTION” is the creation of published ... the field of addiction and homeless services, including more than a decade of ...
(Date:9/20/2017)... ... ... “Psalms of Humidity”: is a fascinating depiction of daily struggles and the ... closer to God. “Psalms of Humidity” is the creation of published author, David Waldrop, ... his life are the very same things that have shaped him into the man ...
(Date:9/20/2017)... (PRWEB) , ... September 20, 2017 , ... “Monique”: ... into uncomfortable situations. “Monique” is the creation of published author, Colleen Crispi, has owned ... Since then Crispi has been involved in real estate and cooking. , “The ...
Breaking Medicine News(10 mins):
(Date:9/1/2017)... Michael Penna , President and CEO ... for growth in his response to the July 13, ... seeking a buyer for eMDs. Penna,s company, Complete HealthCare ... Value Added Reseller and national leader in the independent ... "As the healthcare market continues to dictate consolidation, healthcare ...
(Date:8/31/2017)... 31, 2017 PM360,s annual Innovations Issue, ... the latest innovations happening across the industry. Established six ... focus on providing a comprehensive look at the newest ... the most innovative companies, startups, divisions, products, services, and ... "Everyone in this industry wants to do ...
(Date:8/28/2017)... 2017 Hill-Rom Holdings, Inc. (NYSE: HRC), will present ... 11, 2017, in New York . ... scheduled to present at 11:05 a.m. Eastern Time. ... http://ir.hill-rom.com/events.cfm . A recorded replay will be available one hour ... 2017. About Hill-Rom ...
Breaking Medicine Technology: