NANOPORE SENSING OF AN ANTHRAX PROTIEN Kelby Peterson Mentors: Joseph Robertson & John Suehle Society of Physics Students Summer Internship NIST Gaithersburg,

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NANOPORE SENSING OF AN ANTHRAX PROTIEN Kelby Peterson Mentors: Joseph Robertson & John Suehle Society of Physics Students Summer Internship NIST Gaithersburg, MA

Nanopore Sensing F. Patolsky, C.M. Lieber et al. A. Noy, et al. Wilner & Katz eds. QTL Biosensor AP/WIDE WORLD PHOTOS NIST conceived device: future development blood-test-for-lung-cancer/31800/ BiodefenseClinical DiagnosticsTechniques

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, (1997).

Protective Antigen- Anthrax Protein Nguyen, T.L. Three-dimensional model of the pore form of anthrax protective antigen. Structure and biological implications. J Biomol Struct Dyn 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, (2008).

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

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

Experimental Setup

Experimental System

Rate of Insertion Time (sec) Current (pA)

Rate of Insertion Time (sec) Current (pA)

Understanding Channel Insertion

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

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

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

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