Protein_Sequencing

Card Set Information

Author:
lukemlj
ID:
104114
Filename:
Protein_Sequencing
Updated:
2011-09-25 02:38:46
Tags:
Protein Sequencing
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Description:
Protein Sequencing
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  1. Step 1: Complete Hydrolysis
    HCl 6.0M, 110 C, 16-72 Hrs
  2. Edman Degredation
    • Determine 1st residue (Nth terminal)
    • pH 9.0
  3. Dansylation
    • Determine 1st residue (Nth terminal)
    • Less harsh, faster than Edman
  4. 2-ME or DTT
    Cleave disulfide bond
  5. Iodoacetate
    Prevent formation of disulfide bond
  6. CNBr
    Cleaves on C-side of methionine
  7. Trypsine
    Cleaves lys, arg on C-side
  8. Chemotrypsin
    Cleaves phe, tyr, try on C-side
  9. Endopeptidase S. Aureus V8
    Cleaves asp, glu on C-Side
  10. Elastase
    Cleaves gly, ala, val, syr on C-side
  11. Carboxypeptide A
    • C-side of all except lys, arg, pro
    • Will not work if R(n-1) = pro
  12. Carboxypeptide B
    • C-side of all
    • Will not work if R(n-1) = pro
  13. Carboxypeptide C
    • Cleaves all on C-side
    • Requires acidic conditions (pH 3.5)
  14. Carboxypeptide Y
    • Cleaves all on C-side
    • Slow with gly
  15. Pepsin
    • Cleaves leu, phe, trp, tyr on N-side
    • Will not work if R(n-1) = pro
  16. UV-Absorbance @ 280nm
    Phe, tyr, trp
  17. Fluorescence
    trp

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