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Unravelling the Working of the Animal Body: A Biophysical Approach David A D Parry Institute of Fundamental Sciences Masse...
<ul><li>Replication - DNA </li></ul><ul><li>Protection and Shape – Hair and Skin    (and Bone) </li></ul><ul><li>Locomotio...
Replication
Replication X-ray pattern of DNA Maurice Wilkins Double-Helical Structure
Protection and Shape
Wool and Hair
Hair Follicle <ul><li>Synthesis and assembly of trichocyte keratin molecules into IF occurs in a reducing environment in t...
Molecular Structure of the Hair  Keratin Molecule
Crosslinking Methodology <ul><li>Cross-linking juxtaposed lysine residues of intact IF or sub-assemblies of them with the ...
Positions of the Characterised Crosslinks in Trichocyte Keratin
Axial Parameters Defining IF Structure K1/5/10/14 Reduced   -keratin Oxidised   -keratin No linkers fixed L1, L12 and L2...
Hair Keratin STEM Data 32 chains in cross-section
Key Physical Parameters in Hair Keratin Reduced 3.5 nm/wet/cryo-em 3.5 nm/wet/X-rays 44.92 nm/wet/crosslinks Oxidised 3.0 ...
Models for the “Reduced” and “Oxidised” Hair Keratin IF Reduced (8 + 0) Oxidised (7 + 1)
Locomotion
Components of Connective Tissues  in Mature Animals
Sequence of Collagen Type I Chain
Interaction Maxima Between Parallel Molecules
Molecular Arrangement in Collagen Fibrils
Passive and Active Skins Greyhound Skin Guinea Pig Footpad Skin Trout Skin
Cross-sections of Common Digital Extensor Tendon top) and Suspensory Ligament (Bottom) from  a 5 Year Old Horse
Collagen Fibril Diameter Distributions in Horse Tendon and Ligament
X-ray Picture of Relaxed and Contracting Muscle a  b Low-angle X-ray diffraction patterns from ( a ) relaxed and ( b ) act...
Cross-section Of Muscle Thin Filament Calculated X-Ray Intensities on the Second and Third Layer Lines of the Thin Filament
Regulation Model for Vertebrate Skeletal Muscle
Actin Filament Structure Actin plus tropomyosin structure determined by electron microscopy of isolated actin filaments  i...
Vision
Ultrastructure of Cornea
Corneal Fibril Diameters
Ultrastructure of Cornea and GAGs Parfitt et al. 2010 J. Struct. Biol., 170, 392-397
Corneal Transparency <ul><li>Experimental </li></ul><ul><li>Constant diameter fibrils and a defined range of nearest neghb...
<ul><li>If the separation between fibrils <λ/2 the scattered light from each fibril will interfere destructively in all bu...
Unraveling the Working of the Animal Body: A Biophysical Approach (From left to right) Andrew Miller, David Parry, Barbara...
David Hulmes Alan Craig John Squire Carolyn Cohen Tammy Lynch Peter Steinert
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9.15 k1 d parry

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Plenary 1: D Parry

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9.15 k1 d parry

  1. 1. Unravelling the Working of the Animal Body: A Biophysical Approach David A D Parry Institute of Fundamental Sciences Massey University New Zealand
  2. 2. <ul><li>Replication - DNA </li></ul><ul><li>Protection and Shape – Hair and Skin (and Bone) </li></ul><ul><li>Locomotion – Tendon and Muscle </li></ul><ul><li>Vision - Cornea </li></ul>The Role of Fibrous Molecules
  3. 3. Replication
  4. 4. Replication X-ray pattern of DNA Maurice Wilkins Double-Helical Structure
  5. 5. Protection and Shape
  6. 6. Wool and Hair
  7. 7. Hair Follicle <ul><li>Synthesis and assembly of trichocyte keratin molecules into IF occurs in a reducing environment in the column of cortical cells immediately above the dermal papilla. </li></ul><ul><li>IFAP are deposited and interdigitate with the IF at a later stage towards the end of IF synthesis. </li></ul><ul><li>Further up the hair follicle during terminal differentiation and cell death the cellular environment changes to an oxidising one and disulphide bond formation occurs. </li></ul>
  8. 8. Molecular Structure of the Hair Keratin Molecule
  9. 9. Crosslinking Methodology <ul><li>Cross-linking juxtaposed lysine residues of intact IF or sub-assemblies of them with the periodate-cleavable bifunctional cross-linking reagent DST under mild conditions that do not prevent subsequent IF assembly of the modified proteins. </li></ul><ul><li>Cleaving the cross-linked IF proteins with CNBr and trypsin, and resolving the peptides by HPLC. </li></ul><ul><li>Comparing the peptide peaks before and after cross-linking to reveal shifted peaks that could then be recovered for protein chemical characterisation. </li></ul><ul><li>Reacting with periodate to reveal two peptide peaks that could be used for sequencing, thus identifying the lysine residues that had been adjoined by the cross-linker. </li></ul>
  10. 10. Positions of the Characterised Crosslinks in Trichocyte Keratin
  11. 11. Axial Parameters Defining IF Structure K1/5/10/14 Reduced  -keratin Oxidised  -keratin No linkers fixed L1, L12 and L2 Fixed z a 79.74 78.15 50.00 z b 110.71 112.18 133.30 Repeat 301.16 302.51 316.61 Overlap 7.03 5.67 -8.43 A 12 -0.70 3.30 6.98 A 11 -111.41 -108.88 -126.32 A 22 189.75 193.63 190.28 L1 14.84 14.84 14.84 L12 12.57 12.57 12.57 L2 4.77 4.77 4.77 Δz(1BU, 1BD) -3.91 -1.38 -18.82 Δz(2BU, 2BD) 2.56 6.45 3.10 Δz(1BU, 2D) -50.54 -46.54 -42.86
  12. 12. Hair Keratin STEM Data 32 chains in cross-section
  13. 13. Key Physical Parameters in Hair Keratin Reduced 3.5 nm/wet/cryo-em 3.5 nm/wet/X-rays 44.92 nm/wet/crosslinks Oxidised 3.0 nm/wet/X-rays 2.9 nm /dry/X-rays 47.0 nm/wet/X-rays Radius of Electron-dense ring Axial Repeat r (8 + 0) r (7 + 1) Protofibrils contain four chains (two molecules) in cross-section
  14. 14. Models for the “Reduced” and “Oxidised” Hair Keratin IF Reduced (8 + 0) Oxidised (7 + 1)
  15. 15. Locomotion
  16. 16. Components of Connective Tissues in Mature Animals
  17. 17. Sequence of Collagen Type I Chain
  18. 18. Interaction Maxima Between Parallel Molecules
  19. 19. Molecular Arrangement in Collagen Fibrils
  20. 20. Passive and Active Skins Greyhound Skin Guinea Pig Footpad Skin Trout Skin
  21. 21. Cross-sections of Common Digital Extensor Tendon top) and Suspensory Ligament (Bottom) from a 5 Year Old Horse
  22. 22. Collagen Fibril Diameter Distributions in Horse Tendon and Ligament
  23. 23. X-ray Picture of Relaxed and Contracting Muscle a b Low-angle X-ray diffraction patterns from ( a ) relaxed and ( b ) active molluscan smooth muscle (anterior byssus retractor muscle from Mytilus edulis ). The actin pattern in ( a ) shows a clear third layer line but a missing second layer line, whereas ( b ) shows a second layer line (2) almost as strong as the third (3). From Vibert et al. [3].
  24. 24. Cross-section Of Muscle Thin Filament Calculated X-Ray Intensities on the Second and Third Layer Lines of the Thin Filament
  25. 25. Regulation Model for Vertebrate Skeletal Muscle
  26. 26. Actin Filament Structure Actin plus tropomyosin structure determined by electron microscopy of isolated actin filaments in the on and off states. Reconstructions by Lehman, Craig,Vibert et al.
  27. 27. Vision
  28. 28. Ultrastructure of Cornea
  29. 29. Corneal Fibril Diameters
  30. 30. Ultrastructure of Cornea and GAGs Parfitt et al. 2010 J. Struct. Biol., 170, 392-397
  31. 31. Corneal Transparency <ul><li>Experimental </li></ul><ul><li>Constant diameter fibrils and a defined range of nearest neghbour separations. </li></ul><ul><li>Theoretical </li></ul><ul><li>Light impinging on collagen fibrils is scattered in all directions but as the fibrils all have the same diameter the scattered light has equal intensity and wavelength. </li></ul>
  32. 32. <ul><li>If the separation between fibrils <λ/2 the scattered light from each fibril will interfere destructively in all but the forward direction  light entering the cornea will pass through unaffected. </li></ul><ul><li>For non-uniform distributions of fibril size and random (larger) fibril separations the scattered light will interfere destructively making the cornea opaque. </li></ul>Corneal Transparency
  33. 33. Unraveling the Working of the Animal Body: A Biophysical Approach (From left to right) Andrew Miller, David Parry, Barbara Brodsky, Bruce Fraser, Tom MacRae, Eikichi Suzuki
  34. 34. David Hulmes Alan Craig John Squire Carolyn Cohen Tammy Lynch Peter Steinert

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