determine protein sequence

determine protein sequence

1. aa composition: acid hydrolysis

   • 6M HCL 10-100
     • acid destroy Trp, Gln Asn--> Glu, Asp+NH+; Gln+glu, and Asn+Asp were determined.
   • chromatograpny
     • ion exchange

2. FDNB nerinatization of amino terminal

   • DNP-aa
     • N-term Ala/ internal Lys/ N-term Lys PSQs
       • phase, partition, ether, water, Interlys: water; n-term: ether

3. edman deragation

   • PITC
     • high ph, unportonated, Nu: attack PITC
       • chromato
         • purification

4. PTH aa

   • hydrophobic stationary phase, polar mobile phase

5. Cyanogen bromide

   • COOH side of Met, lactone at new COOH terminals
     • XXXXXMcooh, XXXXMcooh,

6. Trypsin, chymotrypsin, cleavage C terminal

   • trypsin: Arg,lys
     • XXXRcoo, XXXKcoo
   • Chymotrypsin: phe, tyr, trp

7. L7-18