Covalent structure of Proteins or Primary structure of protein:

Proteins are the work horse of biochemical machinary in living systems. They perform various functions from catalyzing the reactions (enzymes), structural (karetin, chitin etc), immunological defense to ion channels. The functional diversity of proteins are due to the difference in their primary, secondary, tertiary and quaternary structures.

Primary structure: Amino acid sequence of polypeptide chain. Secondary Structure: The spatial arrangement of amino acid residue with regard to the back bone structure of polypeptide. Tertiary Structure: Three dimensional structure of entire peptide including the folding of peptide chains. Quaternary structure: When more than one polypeptide units are assembled to gather to give the final structure of a protein, the arrangement of different subunits in space is referred as quaternary structure

Primary structure of Insulin

Once a protein has been purified and characterized in terms of it molecular mass and pI, the next steps are to find our its amino acid sequence and 2o and 3o structures to study the structure function relationships.
Following analysis are commonly performed to get the primary structure of any protein; 1. Determination of the amino-terminal amino acid residue 2. Determination of Amino acid composition 3. Breaking the disulfide bonds between the peptide chains 4. Digestion of polypeptide into small fragments with different enzymes or reagent which cleave at specific residues 5. Determination of sequence of amino acids in each peptide by Edman degradation 6. Recently mass spectrometric methods have been developed to identify proteins by peptide mass finger printing and peptide sequencing.

Chemical synthesis of peptides: Some peptides are are very important pharmacological reagents and are commercially valuable. Smaller peptides are used extensively for the research purposes such as study of active sites of enzymes or inhibitors, receptor blockers.

Advances and creative approaches in organic chemistry have made it possible to synthesize smaller peptides within hours and days. Merrifields method for solid phase synthesis: The first amino acid on carboxyl end of the peptide is attached to a solid resin and the different amino acids are added one by one to form peptide bond . At the end of the synthesis the peptide is detached from the resin and purified.

Amino acid sequence of human cytochrome c