Chemical Bioconjugation of Proteins in an Undergraduate Lab: One-Pot Oxidation and Derivatization of the N-Terminus

Nicholas David James Yates; Robin Louis Brabham; Richard James Spears; Tessa Keenan; Philip A. Helliwell; David Stephen Pugh; Alison Parkin; Glenn A. Hurst; Martin Anthony Fascione

doi:10.1021/acs.jchemed.8b00787

Chemical Bioconjugation of Proteins in an Undergraduate Lab: One-Pot Oxidation and Derivatization of the N-Terminus

Nicholas David James Yates, Robin Louis Brabham, Richard James Spears, Tessa Keenan, Philip A. Helliwell, David Stephen Pugh, Alison Parkin, Glenn A. Hurst, Martin Anthony Fascione

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

A laboratory experiment introducing the concept of chemical bioconjugation of proteins to undergraduate students in a therapeutically relevant context was developed. Initially, students installed an aldehyde functionality into a protein via the oxidation of the N-terminal threonine residue of the cholera toxin subunit B (CTB) protein, which was followed by subsequent modification via hydrazone addition under mild conditions with a chromophore bearing a distinct UV–vis-absorption peak. Students determined the yield of the reaction to be ca. 11% by HPLC coupled to UV–vis spectroscopy and developed key skills such as preparation of stock solutions, chemical manipulation of proteins, and analysis via HPLC. The reported experiment can be readily adapted for use with other proteins and may contribute to enhancing constructive alignment in interdisciplinary degree programs at the chemistry–biology interface.

Original language	English
Number of pages	5
Journal	JOURNAL OF CHEMICAL EDUCATION
DOIs	https://doi.org/10.1021/acs.jchemed.8b00787
Publication status	Published - 26 Mar 2019

Bibliographical note

© 2019 American Chemical Society and Division of Chemical Education, Inc. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details.

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10.1021/acs.jchemed.8b00787

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© 2019 American Chemical Society and Division of Chemical Education, Inc. This is an author-produced version of the published paper. Uploaded in accordance with the publisher’s self-archiving policy. Further copying may not be permitted; contact the publisher for details.
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Cite this

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title = "Chemical Bioconjugation of Proteins in an Undergraduate Lab: One-Pot Oxidation and Derivatization of the N-Terminus",

abstract = "A laboratory experiment introducing the concept of chemical bioconjugation of proteins to undergraduate students in a therapeutically relevant context was developed. Initially, students installed an aldehyde functionality into a protein via the oxidation of the N-terminal threonine residue of the cholera toxin subunit B (CTB) protein, which was followed by subsequent modification via hydrazone addition under mild conditions with a chromophore bearing a distinct UV–vis-absorption peak. Students determined the yield of the reaction to be ca. 11% by HPLC coupled to UV–vis spectroscopy and developed key skills such as preparation of stock solutions, chemical manipulation of proteins, and analysis via HPLC. The reported experiment can be readily adapted for use with other proteins and may contribute to enhancing constructive alignment in interdisciplinary degree programs at the chemistry–biology interface.",

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AU - Yates, Nicholas David James

AU - Brabham, Robin Louis

AU - Spears, Richard James

AU - Keenan, Tessa

AU - Helliwell, Philip A.

AU - Pugh, David Stephen

AU - Parkin, Alison

AU - Hurst, Glenn A.

AU - Fascione, Martin Anthony

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