Protein Sequencing Flashcards
(36 cards)
Step 1 of Protein Sequencing
Hydrolyze to constituent amino acids/ Determination of amino acid concentration
Step 2 of Protein Sequencing
Determination of N-terminal and C-terminal amino acids
Step 3 of Protein Sequencing
Fragmentation of polypeptide chain/ Cleave protein
Step 4 of Protein Sequencing
Reconstruction of amino acid sequence
Protein Hydrolysis
Protein or smaller peptide is strong acid or base solution is heated/ Reverse of the formation of peptide bond reaction
Protein Hydrolysis
Protein or smaller peptide is strong acid or base solution is heated/ Reverse of the formation of peptide bond reaction
Product of protein hydrolysis
free amino acids
Protein Digestion
Enzyme-catalyzed hydrolysis of ingested proteins
Protein Digestion scenario
free amino acids > intestinal wall > bloodstream > liver > raw material for protein synthesis
Chemicals for determination of N-terminal
-Phenylisothiocyanate
-2,4-dinitrofluorobenzene (DNFB)
-Dansyl Chloride
Edman degradation uses what chemical
phenyl isothiocyanate
Edman Degradation products
5-substituted N-phenylthiohydantoin and shorter peptide
Determination of C-terminal;
all except pro, arg, lys
Carboxypeptidase A
Determination of C-terminal;
only for arg and lys
Carboxypeptidase B
Determination of C-terminal;
any residue
Carboxypeptidase C or Y
Carboxypeptidase A
All except pro, arg, lys
Carboxypeptidate A
All except pro, arg, lys
Carboxypeptidase B
only arg and lys
Carboxypeptidase C
Any residue
Carboxypeptidase Y
Any residue
Trypsin
C-terminal of arg, lys
C-terminal of arg, lys
Trypsin
Chymotripsin
C-terminal of phe, tyr, trp
C-terminal of phe, tyr, trp
Chymotrypsin
