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Molecular probes identify changes in fibronectin that may lead to disease
Date:4/24/2012

cular events occur in living organisms has not yet been presented, according to Barker.

A technique called intramolecular fluorescence resonance energy transfer (FRET) has been used to detect molecular strain events in fibronectin fibers, but the technique has limitations because it cannot be used on living tissues and requires the fibronectin to be chemically labeled.

"The molecular probes we identified can be used to map molecular strain events in native extracellular matrix and living lung tissues," explained Barker. "The probes can also be used to study the mechanism by which cells control the mechanical forces that alter fibronectin's conformation, control the exposure of its binding sites and regulate cell signaling."

The researchers used a controlled fibronectin fiber deposition and extension technique to apply tension to the fibers and stretch them to 2.6 times their original length without significant breakage. Then they used a technique called phage display to identify peptides capable of discriminating fibronectin fibers under relaxed and strained conditions. The molecular probes displaying peptide sequences LNLPHG and RFSAFY showed the greatest binding affinity to fibronectin fibers and the greatest efficiency in discriminating between relaxed and strained fibers.

For proof-of-concept demonstrations, the researchers used the probes to discriminate fibronectin fibers within native extracellular matrix and mouse lung slices. LNLPHG preferentially attached to relaxed fibronectin fibers, whereas RFSAFY bound to strained fibers. The probes never attached to the same fiber, which confirmed their ability to selectively discriminate regions within a fibronectin fiber network.

"This study strongly suggests that fibronectin fibers under strain display markedly different biochemical signatures that can be used for the molecular-level detection of fibronectin fiber strain," explained Barker. "The data also show the poten
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Contact: Abby Robinson
abby@innovate.gatech.edu
404-385-3364
Georgia Institute of Technology Research News
Source:Eurekalert  

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