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What reaction is catalyzed by proteases? What steps are
involved in determining the primary structure of a protein?
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Answer Posted / rajesh
A protease is any enzyme that conducts proteolysis; that is,
begins protein catabolism by hydrolysis of the peptide bonds
that link amino acids together in the polypeptide chain.
Classification
Proteases are currently classified into six groups:
* Serine proteases
* Threonine proteases
* Cysteine proteases
* Aspartic acid proteases (e. g., plasmepsin)
* Metalloproteases
* Glutamic acid proteases
steps in determining the primary protein:
1.obtain a pure protein
2.determine the aminoacid composition and M.Wt of
proteins,from this calculate no. of aminoacid residues
present per protein molecule to the nearest whole number.
3.Reduce disulfide groups to sulfhydryl groups.
4.determine N-terminal and c-terminal.
5.fragment aliquots of polypeptide by enzymatic or chemical
hydrolysis and seperate peptide mixtures into individual
peptide fragments.
6.sequence each fragment,and by aanalysing the overlapping
parts,assemble the sequence of original protein.
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