BIOL-2070H_LAB-3&4-WRITEUP.docx - 1 Investigation of...

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1Investigation of Erythrocyte MembraneProteins in Bovinae using SDS-PAGE
2Abstract:Proteins are an essential aspect of cells that enable biological and physiological functions to exist and be carried out in most living organisms. Moreover, proteins found in erythrocytes, or red blood cells, may have a multitude of properties, based on their unique architecture and size. In this lab, the main goal was utilize SDS-PAGE to allow usto separate and identify proteins from a sample of bovine blood and determine if these proteins are also present in Homo sapiens. To do this, we worked with retention factors and molecular weights of these unidentified proteins. The three identified proteins had molecular weights of 117.8 kDa, 29.7 kDa, and 22.8 kDa, and could be identified as Ras GTPase-activating protein 1, Aquaporin-1, and Ras-related protein Rap-2a. Since these proteins are found in the erythrocytes of both bovinae and Homo sapiens, support for the hypothesis was given. Furthermore, our findings allowed us to evaluate the overall effectiveness of SDS-PAGE.Introduction:The molecular structure of cellular membranes has been an important aspect of study in cellular biology, as structure is very closely associated to function. Contained within these membranes are different proteins that perform specialized tasks based on their composition. Another contributor to protein function is the cell’s strong, yet fluid-like nature (Alberts et al. 2014). In erythrocytes, this property is what enables the cells to transport in and out of small capillaries without disrupting them. This is all made possibleby the internal arrangement of spectrin proteins (Kakhniashvili et al. 2004). However, more information about membrane proteins, such as their organization is yet to be
3explored and evaluated. In order to do so, these proteins can be subjected to identificationvia sodium dodecylsulphate-polyacrylamide gel electrophoresis, or SDS-PAGE, which separates the individual polypeptides within. In this process, sodium dodecylsulphate detergent is added to the proteins and placed in a polyacrylamide gel matrix. After, the mixture is exposed to an electrical field, where the attractive cathode and repellant anode enable the separated protein polypeptides to be distinguished according to the size differences. By applying a Coomasie Brilliant Blue dye to stain, the protein bands can be compared to known standard protein molecular weights and identified (Dew and Kapron 2016). The main focus of this study is to determine whether or not the unidentified proteins in the plasma membrane of bovine erythrocytes bears resemblance to that of Homo sapiens. By distinguishing the protein bands based on their various molecular weights and corresponding functions, the proteins can be identified (Caprette 2012).

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