Metal Dyshomeostasis in Alzheimer’s Disease: An Observation of the Role of ZIP12 in Zinc Uptake in Neuronal Cells

Brown, Madison; Whitney, Morgan; Lancellotti, Lily; Walker, Allison

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Metal Dyshomeostasis in Alzheimer’s Disease: An Observation of the Role of ZIP12 in Zinc Uptake in Neuronal Cells

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Zinc (Zn²⁺) is a central component in brain development and function. Dyshomeostasis of Zn²⁺ can result in numerous diseases. This paper focuses on ZIP12, a Zn²⁺ transporter protein that facilitates the uptake of Zn²⁺ into cells. ZIP12 plays a crucial role in various cellular functions and ensures that Zn²⁺ is adequately present in the cell to regulate neuronal plasticity, cell growth, and cellular function. This project aimed to mutate ZIP12, which was introduced into the genome of SH-SY5Y cells, to mimic disease associated missense mutations and examine the effect these alterations would have on Zn²⁺ uptake. The mutations, H551N and H584R, were found using the National Institutes of Health (NIH) database and were both present in highly conserved regions of the transmembrane domain of the ZIP12 protein. The hypothesis was that altering a highly conserved residue in the transmembrane domain would diminish the function of ZIP12 by slowing Zn²⁺ uptake. Thermal cycling, digestion, transformation, and isolation of the DNA were used to introduce the mutation into competent E. coli cells. Following sequencing and gel electrophoresis, SH-SY5Y cells were transfected with the mutated DNA. To test the hypothesis, the Zn²⁺ uptake, Vmax, and Km were determined using atomic absorption spectroscopy, and normalized with a BCA assay. H584R had a lower Vmax and higher Km than that of the wild-type (WT), signifying a decreased functional ability of the protein. The findings of this paper are consistent with the original hypothesis and may provide guidance for future studies involving metal uptake into neuronal cells and potential causes of neurological diseases.

This report represents the work of one or more WPI undergraduate students submitted to the faculty as evidence of completion of a degree requirement. WPI routinely publishes these reports on its website without editorial or peer review.

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