NANOPORE SENSING OF AN ANTHRAX PROTIEN Nanopore Sensing Wilner - - PowerPoint PPT Presentation

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NANOPORE SENSING OF AN ANTHRAX PROTIEN Nanopore Sensing Wilner - - PowerPoint PPT Presentation

Sep 01, 2023 22 likes •174 views

Kelby Peterson Mentors: Joseph Robertson & John Suehle Society of Physics Students Summer Internship NIST Gaithersburg, MA NANOPORE SENSING OF AN ANTHRAX PROTIEN Nanopore Sensing Wilner & Katz eds. http://www.biomedicalblog.com/a-

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SLIDE 1

NANOPORE SENSING OF AN ANTHRAX PROTIEN

Kelby Peterson Mentors: Joseph Robertson & John Suehle Society of Physics Students Summer Internship NIST Gaithersburg, MA

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SLIDE 2

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

http://www.biomedicalblog.com/a- blood-test-for-lung-cancer/31800/

Biodefense Clinical Diagnostics Techniques

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SLIDE 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).

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SLIDE 4

Protective Antigen- Anthrax Protein

Nguyen, T.L. Three-dimensional model

  • f 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

  • f the anthrax toxin pore. Nat Struct. Mol. Biol. 15, 754-760 (2008).
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SLIDE 5

Neutron Reflectometry

McGillivray, Valincius, Heinrich, Robertson, Vanderah, Febo-Ayala, Ignatjev, Lösche, and Kasianowicz, 2009. Biophys. J. 96, 1547.

z-uncertainty = 1.1 Å

McGillivray, Valincius, Heinrich, Robertson, Vanderah, Febo-Ayala, Ignatjev, Lösche, and Kasianowicz, 2009. Biophys. J. 96, 1547.

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SLIDE 6

Optimizing Pore Formation

Milne, J.C., et al. Anthrax protective antigen forms

  • ligomers during intoxication of mammalian cells. J Biol

Chem 269, 20607–20612 (1994). Michaelman-Ribero et al.

Aggregation dominates Little pore insertion Best so far

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SLIDE 7

Experimental Setup

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SLIDE 8

Experimental System

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SLIDE 9

Rate of Insertion

Time (sec) Current (pA)

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SLIDE 10

Rate of Insertion

Time (sec) Current (pA)

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SLIDE 11

Understanding Channel Insertion

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SLIDE 12

Special Thanks

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

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SLIDE 13

Summary

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

Difficulties with the System

With FOS-14 Detergent Without FOS-14 Detergent

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SLIDE 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