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Nanopore Sensing of An Anthrax Protien

Nanopore Sensing of An Anthrax Protien. Kelby Peterson Mentors: Joseph Robertson & John Suehle Society of Physics Students Summer Internship NIST Gaithersburg, MA. Nanopore Sensing. Wilner & Katz eds. http://www.biomedicalblog.com/a-blood-test-for-lung-cancer/31800/.

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Nanopore Sensing of An Anthrax Protien

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  1. NanoporeSensing of An Anthrax Protien Kelby Peterson Mentors: Joseph Robertson & John Suehle Society of Physics Students Summer Internship NIST Gaithersburg, MA

  2. Nanopore Sensing • Wilner & Katz eds. • http://www.biomedicalblog.com/a-blood-test-for-lung-cancer/31800/ • AP/WIDE WORLD PHOTOS • A. Noy, et al. • NIST conceived device: future development • F. Patolsky, C.M. Lieber et al. • QTL Biosensor Techniques Biodefense Clinical Diagnostics

  3. Understanding Anthrax Anthrax toxin • 3 Proteins • Protective Antigen • Edema Factor (EF) • Lethal Factor (LF) Petosa, C., et al. Crystal structure of the anthrax toxin protective antigen. Nature 385, 833-838 (1997).

  4. Protective Antigen- Anthrax Protein • Nguyen, T.L. Three-dimensional model of the pore form of anthrax protective antigen. Structure and biological implications. J BiomolStructDyn 22, 253–265 (2004). Katayama, h. et al. GroEL as a molecular scaffold for structural analysis of the anthrax toxin pore. Nat Struct. Mol. Biol. 15, 754-760 (2008).

  5. Neutron Reflectometry • z-uncertainty = 1.1 Å • McGillivray, Valincius, Heinrich, Robertson, Vanderah, Febo-Ayala, Ignatjev, Lösche, and Kasianowicz, 2009. Biophys. J. 96, 1547. • McGillivray, Valincius, Heinrich, Robertson, Vanderah, Febo-Ayala, Ignatjev, Lösche, and Kasianowicz, 2009. Biophys. J. 96, 1547.

  6. Optimizing Pore Formation Aggregation dominates Little pore insertion Best so far • Milne, J.C., et al. Anthrax protective antigen forms oligomers during intoxication of mammalian cells. J BiolChem 269, 20607–20612 (1994). • Michaelman-Ribero et al.

  7. Experimental Setup

  8. Experimental System

  9. Rate of Insertion Current (pA) Time (sec)

  10. Rate of Insertion Current (pA) Time (sec)

  11. Understanding Channel Insertion

  12. Special Thanks Joseph Robertson, NIST John Suehle, NIST Toni Sauncy, SPS Kendra Redmond, AIP David Peak, USU

  13. Summary • Understanding Nanopores is useful • Goal: Neutron Reflectometry • Optimize Channel Insertion • FOS-14 Detergent • Repeatability • No increased nanopore insertion

  14. Difficulties with the System With FOS-14 Detergent Without FOS-14 Detergent

  15. Conclusions • Detergent does not inhibit nanopore formation • Detergent decreases variability in measurements • Detergent does not increase max channel formation • Although detergent does not increase the total channel formation it may be useful for increasing repeatability of measurements

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