BCHE 395-6 2013 Protein Motifs and Sequencing

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Unformatted text preview: ino acid is identified through various methods 27 3. N- terminal sequencing Chemical Reac1ons of Amino Acids Reac1ons of α- amino group Phenylisothiocyanate Under mild acidic condi1on will cyclize Used for protein sequencing no color Also called Edman’s reagent Can be quan1fied on columns 28 3. N- terminal sequencing Chemical Reac1ons of Amino Acids: Sanger Reagent Also used for N- termini sequencing of proteins Reac1ons of α- amino group 2. 1- Fluoro- 2,4- dinitrobenzene (FDNB) 29 Methods for I° Structure Determina1on •  4. Pep@de Fragmenta@on: •  Edman degrada1on can reliably be used for only the first six amino acids •  Limi1ng à༎ recovery of shortened pep1de –  ~93% azer each cycle –  (0.93)60 = 0.013 = 1.3% of original star1ng material azer 60 cycles –  If (0.99)120 = 0.30 = 30% of star1ng material •  Methods designed to generate very specific, quan1ta1ve, and reproducible fragmenta1on of proteins by par@al hydrolysis 30 4. Pep1de Fragmenta1on •  Cleave protein into smal...
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