(a) Describe in detail how you will determine the primary structure of protein. (b)You have been given a mixture of lysine, histidine and cysteine. The isoelectric point of the amino acids are as follows: Histidine 7.64 Lysine 9.74 Cystenie 5.02 Show how you will separate the mixture into the pure forms. State and describe any instrument that you will use to separate the components in the mixture.
(a) Describe in detail how you will determine the primary structure of protein.
(b)You have been given a mixture of lysine, histidine and cysteine. The isoelectric point of the amino acids are as follows:
Histidine 7.64
Lysine 9.74
Cystenie 5.02
Show how you will separate the mixture into the pure forms. State and describe any instrument that you will use to separate the components in the mixture.
The primary structure of a protein is generally referred to the sequence of amino acids in the polypeptide chain. Primary structure is held together strongly by peptide bonds that joins two adjacent amino acids.
Determination of primary structure:
1.
Protein is unknown, even the DNA sequence information is not known and hence start from scratch.
2.
Purify the protein and separate the chains (if multimer), also reduce disulfides and block free thils.
3.
Fragment and sequence each chain by enzymatically, with endopeptidase, chemically with cyanogen bromide or physically through collision in MS.
4.
Fragmented peptides are sequenced by Mass spectrometry where the mass of small peptides are calculated individually.
5.
Reassemble the sequence based on overlapping fragments join it end to end.
Step by step
Solved in 4 steps
