title: Exploring protein structure and dynamics through a project-oriented biochemistry laboratory module.
abstract: Here, we present a 10-week project-oriented laboratory module designed to provide a course-based undergraduate research experience in biochemistry that emphasizes the importance of biomolecular structure and dynamics in enzyme function. This module explores the impact of mutagenesis on an important active site loop for a biomedically-relevant human enzyme, protein tyrosine phosphatase 1B (PTP1B). Over the course of the semester students guide their own mutant of PTP1B from conception to characterization in a cost-effective manner and gain exposure to fundamental techniques in biochemistry, including site-directed DNA mutagenesis, bacterial recombinant protein expression, affinity column purification, protein quantitation, SDS-PAGE, and enzyme kinetics. This project-based approach allows an instructor to simulate a research setting and prepare students for productive research beyond the classroom. Potential modifications to expand or contract this module are also provided. © 2017 by The International Union of Biochemistry and Molecular Biology, 45(5):403-410, 2017.
authors.affiliation: Department of Chemistry, Washington College, Chestertown, Maryland, 21620.
authors.forename: James M
authors.initials: JM
authors.lastname: Lipchock
authors.affiliation: Department of Chemistry, Washington College, Chestertown, Maryland, 21620.
authors.forename: Patrick S
authors.initials: PS
authors.lastname: Ginther
authors.affiliation: Department of Chemistry, Washington College, Chestertown, Maryland, 21620.
authors.forename: Bonnie B
authors.initials: BB
authors.lastname: Douglas
authors.affiliation: Department of Chemistry, Washington College, Chestertown, Maryland, 21620.
authors.forename: Kelly E
authors.initials: KE
authors.lastname: Bird
authors.affiliation: Department of Chemistry, Yale University, New Haven, Connecticut, 06511.|Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, 06511.
authors.forename: J
authors.initials: J
authors.lastname: Patrick Loria
mesh_terms: D001671:Biochemistry / Q000193:education*; D006801:Humans; D007753:Laboratories; D011487:Protein Conformation; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1 / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D012106:Research / Q000193:education*; D013334:Students
chemical_list: C516607:PTPN1 protein, human; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1
keywords: Integration of research into undergraduate teaching; enzymes and catalysis; laboratory exercises; protein structure function and action mechanism