This Intro to Biochemistry Lab was designed as a Course-Based Undergraduate Research Experience, during which students researched a human glycoprotein thyroid peroxidase (TPO) that plays a role in thyroid hormone synthesis and can cause thyroid diseases when mutated. Our research lab previously expressed, purified, and monitored the activity of the wild-type version of TPO with moderate success, making it an ideal candidate for students to work with in a laboratory learning setting. Various known but uncharacterized mutants of TPO were chosen for students to work with for learning biochemistry techniques. A series of labs were developed that included site-directed mutagenesis & protein expression in <i>E.Coli </i>JM109 cells, purification via nickel-affinity chromatography, and quantification via UV/Vis spectrophotometry. In addition, guaiacol peroxidase enzyme assays were run with both wild-type and mutant TPO and type I horseradish peroxidase (HRP) to compare enzyme activity.

Characterizing mutations of thyroid peroxidase (TPO) as an introduction to biochemistry course-based undergraduate research experience

ACS Spring Meeting 2021

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