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Detecting Anthrax Proteins At Ultralow Concentrations

Date:
September 1, 2005
Source:
National Institute of Standards and Technology
Summary:
A new laboratory method for quickly detecting active anthrax proteins within an infected blood sample at extremely low levels has been developed by researchers at the National Institute of Standards and Technology (NIST), the U.S. Army Medical Research Institute of Infectious Diseases and the National Cancer Institute. The method takes about an hour to get unambiguous results compared to up to several days to get results with current techniques.

A computer model shows side and top views of two different proteins produced by anthrax bacteria. The green molecule is "protective antigen" (PA), which spontaneously forms pores that penetrate organic membranes such as cell walls. The yellow molecule is "lethal factor (LF)." When a voltage is applied across a membrane studded with PA pores, both positive and negative ions flow through. Once LF binds to the pore, however, current only flows in one direction.
Credit: Image credit: T. Nguyen, National Cancer Institute

A new laboratory method for quickly detectingactive anthrax proteins within an infected blood sample at extremelylow levels has been developed by researchers at the National Instituteof Standards and Technology (NIST), the U.S. Army Medical ResearchInstitute of Infectious Diseases and the National Cancer Institute.

Current detection methods rely on injecting live animals orcell cultures with samples for analysis and require up to several daysbefore results are available. Described* in an upcoming issue of theJournal of Biological Chemistry, the new method produces unambiguousresults in about an hour. The researchers hope the system willultimately be useful in developing fast, reliable ways to diagnoseanthrax infections or to quickly screen large numbers of drugs aspossible therapies for blocking the bacteria's toxic effects.

The method works by detecting changes in current flow whenanthrax proteins are present in a solution. An anthrax proteinironically called "protective antigen" spontaneously formsnanometer-scale pores that penetrate the surface of an organicmembrane. When a voltage is applied across the membrane, positively andnegatively charged ions flow freely in both directions through thepore. When additional anthrax proteins called lethal factor (LF) oredema factor (EF) are present, however, the proteins bind to theoutside of the pore and shut down the flow of ions in one direction.This change in current flow depends on the concentration of theproteins in the solution and can detect amounts as low as 10 picomolar(trillionths of a mole).

"We hope this system will lead to a method for rapidlyscreening agents that inhibit the binding of LF or EF to these pores,"says NIST's lead investigator John Kasianowicz.

Live anthrax antibodies seem to do exactly that. Whenantibodies were present in the test solution and then LF was added, thecurrent flow remained unchanged, indicating that the anthrax proteinswere unable to bind properly. The long-term goal would be to find drugswith few side effects that also interfere with this binding process.

###

* K.M. Halverson, R.G. Panchal, T. Nguyen, R. Gussio, S.F. Little,M. Misakian, S. Bavari and J.J. Kasianowicz, "Anthrax Biosensor:Protective Antigen Ion Channel Asymmetric Blockade," Journal ofBiological Chemistry, slated for a November issue, posted online Aug.8, 2005.


Story Source:

The above story is based on materials provided by National Institute of Standards and Technology. Note: Materials may be edited for content and length.


Cite This Page:

National Institute of Standards and Technology. "Detecting Anthrax Proteins At Ultralow Concentrations." ScienceDaily. ScienceDaily, 1 September 2005. <www.sciencedaily.com/releases/2005/08/050831074045.htm>.
National Institute of Standards and Technology. (2005, September 1). Detecting Anthrax Proteins At Ultralow Concentrations. ScienceDaily. Retrieved August 28, 2014 from www.sciencedaily.com/releases/2005/08/050831074045.htm
National Institute of Standards and Technology. "Detecting Anthrax Proteins At Ultralow Concentrations." ScienceDaily. www.sciencedaily.com/releases/2005/08/050831074045.htm (accessed August 28, 2014).

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