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IGERT Progress Report November 2007

IGERT Progress Report November 2007. Robert Camp Mechanical Engineering Extracellular Matrix Engineering Research Laboratory (EMERL) Advisor: Jeff Ruberti. Agenda. Collagen – The protein of interest Review of Research Literature Review Pipette Manufacture Chemistry

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IGERT Progress Report November 2007

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  1. IGERT Progress ReportNovember 2007 Robert Camp Mechanical Engineering Extracellular Matrix Engineering Research Laboratory (EMERL) Advisor: Jeff Ruberti

  2. Agenda • Collagen – The protein of interest • Review of Research • Literature Review • Pipette Manufacture • Chemistry • Current Experiment Setup • Question/Comments

  3. What is Collagen? • Principal load bearing and most abundant protein in vertebrate animals • Defined as (GLY-X-Y)n • 3 left-handed helical alpha chains form one right-handed helix collagen monomer Picture Source: Wikipedia.org

  4. Collagen Type I • 27 different types of collagen found in nature • Collagen Type I is the basis of research • One of the fibril forming collagens • Type I, II, III, V, XI • Monomer Size • 300 nanometers long • 1.5 nanometers in diameter Picture Source: Wikipedia.org

  5. Review of Research • Reading key published works concerning my research • Single molecule manipulation • Bustamante - Effects of mechanical force on DNA/RNA polymerase activity • Magnetic Tweezers • Prentiss - Studying the forces in ligand-receptor Complexes (basis of new experiments) • Marko - Single DNA strand manipulation (basis of ultimate experiment) • Collagen • Sun – Mechanical strength of collagen

  6. Bead Catching Apparatus • Manufacturing bead catching pipettes • This is extremely difficult since the tips are so small Permanent Magnet Bead Catching Pipette Source: Yan, et al 2004 Loading Pipette Force Measuring Pipette

  7. Chemistry • Shifting the focus of my current research to the chemical bonding of collagen • Working with antibodies provided by the NICDR • Currently setting up new experiment to test collagen • This will be a springboard to my single collagen manipulation experiments • It will also provide much statistical data to help support my single collagen experiment

  8. Current Experiment Collagen Bind 1 Micron Paramagnetic Beads Wash Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y Not to Scale

  9. Apply Force (10-300 pN) Add MMP-8 Bead and Collagen are to scale

  10. Questions/Comments “The most exciting phrase to hear in science, the one that heralds new discoveries, isn’t ‘eureka!’, but rather ‘hmmmm, that’s funny’.” Isaac Asimov

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