Start Date
2023
Description
GluT-1 is the primary glucose transporter protein found in the membrane of mammalian cells. Not only does the type of cell determine the concentration of the transporter protein in the cell’s membrane, but also the health and state of the cell. Cancer cells are commonly known to use high concentration of GluT-1 to their advantage to provide more energy for the rapidly growing cells. Scarlet-GluT-1 Introduction Conclusions and Future Directions References Intracellular Interactions of GluT-1 Trevor Rublein, Ben Scofield, Prof. Arnoys, Calvin University, Grand Rapids, Michigan We would like to thank Prof. Laura Westrate, Mr. Dave Ross, Prof. Lori Keen, and Mr. Nick Boersma for their additional help and mentorship in the lab. We would also like to thank Katarina Woldt, Samuel Steen, Hannah King, and Allison Chen for their help and leadership in the lab as peers. Acknowledgements Sec 23 and Sec 24 The Sec proteins, including Sec23a-b, Sec24a-b, and a few others are involved in vesicle formation in the endoplasmic reticulum. The formation of the complex outside the ER membrane allows for GluT-1 to be transported out and sent to the cellular membrane. To express and purify these proteins, we used a variety of tagging methods. Figure 2: A) The mScarlet3 gene was amplified and was cut along with the GFP-GluT-1 vector to have sticky ends, and then A) D) B) E) MEMO ligated to create a full plasmid. The maroon arrows indicate the position and direction of the primer used to amplify the sequence, and the restriction enzymes sites used are shown. B) The complete vector for mScarlet3. C) PCR products of the mScarlet3 insert and GluT-1 vector were run through 10% agarose gel to determine if the products were the correct size. D) An image under the fluorescent microscope of a Cos7 cell transfected with 200ng of the mScarlet3 GluT-1 plasmid using the Lipofectamine 3000 reagent. E) The crystal structure of mScarlet3. The bright red color is caused by a conjugation of amino acids R groups in the middle of the protein structure. C) mScarlet3-insert 0.5kb 2kb 1kb 4kb 3kb GluT-1 consists of 12 transmembrane α-helices that shape the channel for glucose and a small intracellular peptide tail. Other proteins are able to bind to GluT-1 in a variety of places including the active site, the 13-15 residue peptide tail, and others not identified yet.
Recommended Citation
Rublein, Trevor; Scofield, Ben; and Arnoys, Eric, "Intracellular Interactions of GluT-1" (2023). Summer Research. 40.
https://digitalcommons.calvin.edu/summer_research/2023/Posters/40
Included in
Biology Commons, Cancer Biology Commons, Molecular Biology Commons
Intracellular Interactions of GluT-1
GluT-1 is the primary glucose transporter protein found in the membrane of mammalian cells. Not only does the type of cell determine the concentration of the transporter protein in the cell’s membrane, but also the health and state of the cell. Cancer cells are commonly known to use high concentration of GluT-1 to their advantage to provide more energy for the rapidly growing cells. Scarlet-GluT-1 Introduction Conclusions and Future Directions References Intracellular Interactions of GluT-1 Trevor Rublein, Ben Scofield, Prof. Arnoys, Calvin University, Grand Rapids, Michigan We would like to thank Prof. Laura Westrate, Mr. Dave Ross, Prof. Lori Keen, and Mr. Nick Boersma for their additional help and mentorship in the lab. We would also like to thank Katarina Woldt, Samuel Steen, Hannah King, and Allison Chen for their help and leadership in the lab as peers. Acknowledgements Sec 23 and Sec 24 The Sec proteins, including Sec23a-b, Sec24a-b, and a few others are involved in vesicle formation in the endoplasmic reticulum. The formation of the complex outside the ER membrane allows for GluT-1 to be transported out and sent to the cellular membrane. To express and purify these proteins, we used a variety of tagging methods. Figure 2: A) The mScarlet3 gene was amplified and was cut along with the GFP-GluT-1 vector to have sticky ends, and then A) D) B) E) MEMO ligated to create a full plasmid. The maroon arrows indicate the position and direction of the primer used to amplify the sequence, and the restriction enzymes sites used are shown. B) The complete vector for mScarlet3. C) PCR products of the mScarlet3 insert and GluT-1 vector were run through 10% agarose gel to determine if the products were the correct size. D) An image under the fluorescent microscope of a Cos7 cell transfected with 200ng of the mScarlet3 GluT-1 plasmid using the Lipofectamine 3000 reagent. E) The crystal structure of mScarlet3. The bright red color is caused by a conjugation of amino acids R groups in the middle of the protein structure. C) mScarlet3-insert 0.5kb 2kb 1kb 4kb 3kb GluT-1 consists of 12 transmembrane α-helices that shape the channel for glucose and a small intracellular peptide tail. Other proteins are able to bind to GluT-1 in a variety of places including the active site, the 13-15 residue peptide tail, and others not identified yet.