Exploring protein structure and dynamics through a project-oriented biochemistry laboratory module

Biochem Mol Biol Educ. 2017 Sep;45(5):403-410. doi: 10.1002/bmb.21056. Epub 2017 Mar 13.

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.

Keywords: Integration of research into undergraduate teaching; enzymes and catalysis; laboratory exercises; protein structure function and action mechanism.

MeSH terms

  • Biochemistry / education*
  • Humans
  • Laboratories*
  • Protein Conformation
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1 / chemistry*
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1 / isolation & purification
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1 / metabolism*
  • Research / education*
  • Students

Substances

  • PTPN1 protein, human
  • Protein Tyrosine Phosphatase, Non-Receptor Type 1