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Week 2 - review of chemistry, intro to crystallography
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Week 2 - review of chemistry, intro to crystallography

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  • Structural analysis overview
  • Transcript

    • 1. The Structures of Atoms Found in Organisms
    • 2. Different Types of Covalent Bonds the shared electrons glue the atoms together
    • 3.  
    • 4. Basic Structure of An Amino Acid
    • 5. alanine arginine asparagine aspartate cysteine glutamate glutamine glycine histidine isoleucine leucine lysine methionine phenylalanine histidine serine threonine tryptophan tyrosine valine The Twenty Amino Acids
    • 6. Proteins are Polymers of Amino Acids some proteins in your body have the same amino acid sequence as those found in the chimpanzee
    • 7.  
    • 8. Critical Stages in an X-ray Analysis
      • Cloning, over-expression, and protein purification
      • Crystallization
      • Data collection and processing
      • Interpretation of electron density maps
    • 9. Fundamentals of Protein Crystallization
      • Crystallization is simply achieved by reducing the solubility of the protein under conditions where the stable state becomes the crystalline lattice.
      • The problem is finding those conditions!
      • Must determine the appropriate precipitant and state of the protein for crystallization to occur.
    • 10. Finding Crystallization Conditions
      • This is still a matter of trial and error.
      • There is no guarantee that a given protein can be crystallized.
      • Crystallization screens have been developed that improve the chances of finding suitable conditions.
      • A large number of crystallization screens are commercially available from Hampton Research.
    • 11. Important Parameters in Protein Crystallization
      • The state and purity of the protein
      • Precipitant (type and concentration)
      • pH - usually test in 1/2 pH units
      • Temperature
      • Protein concentration
    • 12. Common Precipitants (Salts)
      • Salts
        • ammonium sulfate
        • ammonium phosphate
        • sodium/potassium phosphate
        • sodium citrate
        • sodium sulfate
        • sodium chloride
        • magnesium sulfate
    • 13. Common Precipitants (Organics )
      • polymeric precipitants
        • poly(ethylene glycol) 6000-8000
        • methylether poly(ethylene glycol) 5000
        • poly(ethylene glycol) 1000-2000
        • poly(ethylene glycol) 400
      • alcohols
        • ethanol, propanol
        • 2-methyl-2,4,-pentane diol (MPD), propanediol
    • 14. Vapor Diffusion
      • Introduced in the late 1960s
      • “ Conventional” sitting drop method was introduced by Bob Bock in 1968 (former Dean of the Graduate School at UW)
      • Hanging drop method was developed in 1971
      • This became the standard method for crystallization by 1972
    • 15. Hanging Drop Method of Vapor Diffusion This is the most commonly used technique for surveying and growing crystals for structural analyses.
    • 16. X-ray Diffraction is the Best Test for Crystallinity
      • Most protein crystals are grown with the goal of determining their structure, thus an x-ray diffraction experiment is considered to be the ultimate proof that a well ordered crystalline lattice has been achieved.
    • 17. Common Table Salt
    • 18. Common Table Sugar
    • 19. C-3 ’ -Methyltransferase
      • Involved in the production of an unusual sugar called tetronitrose
      • Tetronitrose is found in an antitumor and antibacterial agent called tetrocarcin A.
    • 20.  
    • 21. C-3 ’ -Methyltransferase
    • 22. S -Adenosylmethionine
      • Discovered in Italy in 1952
      • Referred to as SAMe in the vitamin supplement stores
      • May be helpful in the treatment of depression, osteoarthritis, fibromyalgia, liver disease, Alzheimer’s Disease
    • 23. Space-Filling
    • 24. Ball-and-Stick
    • 25. Ribbon Representation
    • 26. Electron Density
    • 27. His 181 Glu 224 Tyr 222