Post #5: Chimera Science

Hi everyone! As you might remember from my last post, I successfully expressed the autophagy marker LC3 in HeLa cells, but it did not co-localize with my mutant receptor. Since then, I have been researching different methods that I could potentially use to study the protein aggregation of mutant tc-TRa1. One particular method that I am investigating is the use of chimeric proteins. 

Chimeric proteins are artificially created from the fusion of two different genes that originally coded for two separate proteins. These hybrid peptides have several applications in biotechnology and molecular biology. For example, the TRa1 receptors I have used in my research this summer are chimeric proteins, since they were synthesized by fusing a human TRa1 receptor and a fluorescent tag, green fluorescent protein (GFP), originally isolated from the crystal jelly.

Figure 1. Crystal structure of GFP.

Figure 1. Crystal structure of GFP.

There are two chimeric proteins that have been previously used as aggregation markers: GFP-250 and GFP 170. Both were created by fusing GFP with truncated versions of proteins involved in the transport of cargo to the Golgi apparatus. Normally, the full length proteins are localized to the Golgi, but these truncated variants tend to form aggregates near the microtubule organizing center (MTOC), displaying features of an aggresome. Aggresomes are bodies of protein aggregates that form around the MTOC when the cell is incapable of completely degrading misfolded proteins. My plan this week is to co-transfect the mutant tc-TRa1 with GFP 170 and GFP-250 and observe if there is any co-localization between the receptor and the aggresome markers.

Stay tuned for my results!

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