Beta-2-microglobulin mutations

• In Event: Poster Presentation and Judging for: Biomedical; Cell & Molecular Biology; Chemistry & Biochemistry; Marine Science; Systematics & Evolutionary Biology.
  In Poster Session: Chemistry & Biochemistry Poster Session

Abstract

Beta-2-microglobulin (B2M) is a 99 residue protein that is a part of the Class 1 Major Histocompatibility Complex (MHC). The protein is characterized by a 7-stranded B sandwich stabilized by a single disulfide bond. B2M misfolds to aggregate into amyloid fibrils which has been linked to the disease Dialysis Related Amyloidosis (DRA). To investigate the misfolding process 2M will be mutated at different locations of the protein to test their effect on how the protein folds normally and how it misfolds. There are a lot of mutations that have been seen to cause big differences in the lag and elongation of the protein at or around position 60 such as position 58, 60, and 65. The lag and elongation time of protein folding has been directly tied to the formation of amyloid fibrils. The lag and elongation are two stages of the protein misfolding process. Where lag is a measure of the amount of time it takes in order to initiate the misfolding event and the elongation is the rate of propagation. There are also some interesting positions found towards the end of the protein that have major effects of the lag and elongation of the protein. Position 82 is one such position. By changing specific amino acids in specific places in the protein, valuable information will be obtained on how misfolding is occurring. Mutations were made through the process of Quick Change PCR, PCR purification, transformation, and QIAPrep. The mutations were then confirmed with DNA sequencing.

Author

• Mikayla Rose Gascich, Austin College