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Collagen Research: Literature Review, Pipette Manufacture, Chemistry, and Current Experiment Setup

This progress report provides an overview of collagen research, including literature review, pipette manufacture, chemistry, and the current experiment setup. The report also highlights the importance of collagen and the ongoing efforts to manipulate and study its properties.

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Collagen Research: Literature Review, Pipette Manufacture, Chemistry, and Current Experiment Setup

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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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