Introduction: (summary of content that will be explained in detail by group 1) Amino acids contain an amine functional group as well

n a carboxylic functional group Both attached to the same carbon atom they are known as 2-amino acids or -amino acids There are 20 such 2-amino acids that occur naturally and they are the building blocks of proteins Proteins are large macromolecules made up of chains of 2-amino acids The amino acids bond to each other through condensation reactions that result in polypeptides (term used to refer to 4 or more amino acids that are bonded together) After the condensation reaction, only residue of the amino acid remains which join together in a bond referred to as a peptide bond or an amide link Each protein contains a fixed number of amino acid residues that are connected together in a strict sequence that is referred to as the primary structure of the protein.

Protein Analysis: The 2 methods of protein analysis we will be explaining, namely paper chromatography and electrophoresis, can be used to determine the primary structure of a protein. In both methods the protein first undergoes hydrolysis by hydrochloric acid to release the amino acids that will be used for primary structure identification. Paper Chromatography: is a technique that is used to separate and identify substances that are or can be colored After the hydrolyzed protein releases various amino acids, a small sample of an unknown amino acid is placed at the bottom of the chromatographic paper Several samples of known amino acids can be placed alongside The paper is then placed in a solvent which rises up the paper due to capillary action. As the solvent meets the samples, the different samples of amino acids partition themselves between the solvent and the paper to different extent s and move up the paper at different rates. When the solvent has nearly reached the top, the paper is removed from the tank, dried and sprayed with an organic dye to develop the chromatogram by coloring the acids

The position of the spots of different samples, which becomes clearly visible due to the sprayed paper, can then be compared and the sample can be identified based on how much it has moved in relation to the known amino acid samples. This is movement in the position of the sample is referred to as the retention factor (defined as the ratio between the distance travelled by the sample and the distance travelled by the solvent, denoted as Rf) If samples of known amino acids are not available then the retention factor can be measured and compared to the retention factors of known amino acids in order to identify it The few real life instance where this technique is used, most commonly 2-dimensional chromatography is used which involves using two solvents and rotating the paper 90 in between This is more commonly used because it is possible for 2 acids to have the same retention in one solvent and different in another and to ensure that there is different retention obtained for all samples, 2-dimensional chromatography is used Similar compounds have the tendency to have similar retention hence using 2 solvents in 2-dimensional chromatography gives a better probability of obtaining a different retention value Hence this method is useful for separating complex mixtures of similar compounds which is true in the case of amino acids because they are essentially long chains of similar compounds. Paper chromatography has largely been replaced by thin layer chromatography in todays world. However this technique s still great because it is efficient, requires very small samples and the retention values for many substance are known through testing which makes the few unknown substances relatively easy to identify. The secondary structure of the protein, which describes the way in which the chain of amino acids folds itself due to intermolecular hydrogen bonding, can be confirmed using a process called X-ray crystallography.