Using HPLC-Mass Spectrometry to Teach Proteomics Concepts with Problem-based Techniques

Document Type

Article

Publication Date

8-2010

Publication Source

Biochemistry and Molecular Biology Education

Volume Number

38

Issue Number

4

First Page

242

Last Page

246

Publisher

John Wiley & Sons Inc.

ISSN

1470-8175

Abstract

Practical instruction of proteomics concepts was provided using high-performance liquid chromatography coupled with a mass selective detection system (HPLC-MS) for the analysis of simulated protein digests. The samples were prepared from selected dipeptides in order to facilitate the mass spectral identification. As part of the prelaboratory preparation, students calculated the parent ion patterns of the dipeptides using peptide calculator websites. Following instruction on the use of the HPLC-MS instrument, students analyzed mixtures of the dipeptides and identified the individual dipeptides in the unknowns. In addition, purchased chicken egg white lysozyme alkylated with iodoacetamide and digested with trypsin was analyzed using the same approach. Key tryptic peptides were identified from the HPLC-MS chromatogram with information generated with the FindPept tool. This experiment demonstrates that complex concepts can be taught in the undergraduate biochemistry laboratory using a problem-based approach.

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