Identification of an Unknown Amino Acid by Paper Chromatography and Zone Electrophoresis

Unit 5: Identification of an unknown Amino Acid by Paper Chromatography and Zone Electrophoresis

By: Jennifer Brown

Lab Partners:

Karlotta Bushnell

Courtney White

Jasmine Guidry

Kalyn Bowie

Sonya Wolf

I. Abstract

In this experiment, hemoglobin was identified and separated by using paper chromatography and a sickle cell patient was chose based of the results of gel electrophoresis. In the paper chromatography experiment, there was four known samples, Trytophan, Glutamic acid, Serine, Isoleucine. Based on the results, Glutamic Acid was the unknown hemoglobin when separated by paper chromatography. In using gel electrophoresis, there were three samples from the tested people (1, 3, and 4) and two references (2 and 5) . From the findings of these results, 1 and 4 have sickle cell and 3 is normal because the sickle celled amino acid travel slower than the normal amino acid.

II. Introduction

Proteins are large polymers that are made up of amino acids along with other organic molecules such as carbohydrates and lipids. Due to certain protein deficiencies, certain issues and diseases can occur such as edema, fatty liver infiltration and mental retardation. Because proteins play such a huge role in the body, life would not be able to go on without the presence of proteins. The roles of proteins are many, but just to name a few they have a catalytic, protective, storage and structure roles. Proteins generally are found to contain L- amino acids and S- amino acids. Others are stereoisomers which mean they have R configuration instead of the S. The structure of amino acids consist of a central carbon atom, a carboxyl group, a hydrogen, an amino acid group and an unique side chain. Between each amino acid and the carboxyl group of another amino acid, a peptide bond is formed.



In all, there are twenty different amino acids. Most amino acids are chiral, while glycine is achiral. This means that the structure is asymmetric in such a way that the structure and its mirror image are not superimposable. Achiral is just the opposite in that it superimposable on its mirror image. Hemoglobin is often known as the iron containing pigment of red blood cells. It consists of an iron atom in a porphyrin ring structure. Normal blood hemoglobin contains glutamic acid in its primary sequence of amino acids. In sickle hemoglobin, glutamic acid is replaced by valine. Sickle cell can be seen mostly in the African and African American community and cause much pain within an individual.



In this experiment zone electrophoresis was used to determine the outcome of the person with normal versus sickle cell amino acid. Zone electrophoresis is a technique in which components are separated into zones or bands in a buffer, and stabilized in solid, porous, or any other support medium-eg, filter paper, agar gel, or polyacrylamide gel which is used to separate macromolecules such as DNA, RNA, and proteins. Because each DNA molecule is negatively