Selectivity and stability of N-terminal targeting protein modification chemistries

Lydia J. Barber; Nicholas D.J. Yates; Martin A. Fascione; Alison Parkin; Glyn R. Hemsworth; Paul G. Genever; Christopher D. Spicer

doi:10.1039/d2cb00203e

Selectivity and stability of N-terminal targeting protein modification chemistries

Lydia J. Barber, Nicholas D.J. Yates, Martin A. Fascione, Alison Parkin, Glyn R. Hemsworth, Paul G. Genever, Christopher D. Spicer^*

^*Corresponding author for this work

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

Abstract

Protein N-termini provide uniquely reactive motifs for single site protein modification. Though a number of reactions have been developed to target this site, the selectivity, generality, and stability of the conjugates formed has not been studied. We have therefore undertaken a comprehensive comparative study of the most promising methods for N-terminal protein modification, and find that there is no ‘one size fits all’ approach, necessitating reagent screening for a particular protein or application. Moreover, we observed limited stability in all cases, leading to a need for continued innovation and development in the bioconjugation field.

Original language	English
Number of pages	9
Journal	RSC Chemical Biology
Early online date	17 Nov 2022
DOIs	https://doi.org/10.1039/d2cb00203e
Publication status	E-pub ahead of print - 17 Nov 2022

Bibliographical note

Funding Information:
L. J. B. and C. D. S. acknowledge PhD studentship support from the BBSRC White Rose Mechanistic Biology DTP. This PhD was supported with generous industrial CASE support from Qkine and our thanks go to Dr Marko Hyvönen and Dr Catherine Elton for their support. N. D. J. Y. and M. A. F. acknowledge support from the Engineering and Physical Sciences Research Council (EPSRC, EP/V044303/1). Dr Alex Heyam is thanked for help running NMR kinetics experiments. We thank Dr Ed Bergstrom and The York Centre of Excellence in Mass Spectrometry for support with protein mass spectrometry. This centre was created thanks to a major capital investment through Science City York, supported by Yorkshire Forward with funds from the Northern Way Initiative, and subsequent support from the EPSRC (EP/K039660/1; EP/M028127/1).

Publisher Copyright:
© 2022 The Author(s).

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d2cb00203e
© 2022 The Author(s).
Final published version, 5.37 MBLicence: CC BY

Cite this

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title = "Selectivity and stability of N-terminal targeting protein modification chemistries",

abstract = "Protein N-termini provide uniquely reactive motifs for single site protein modification. Though a number of reactions have been developed to target this site, the selectivity, generality, and stability of the conjugates formed has not been studied. We have therefore undertaken a comprehensive comparative study of the most promising methods for N-terminal protein modification, and find that there is no {\textquoteleft}one size fits all{\textquoteright} approach, necessitating reagent screening for a particular protein or application. Moreover, we observed limited stability in all cases, leading to a need for continued innovation and development in the bioconjugation field.",

author = "Barber, {Lydia J.} and Yates, {Nicholas D.J.} and Fascione, {Martin A.} and Alison Parkin and Hemsworth, {Glyn R.} and Genever, {Paul G.} and Spicer, {Christopher D.}",

note = "Funding Information: L. J. B. and C. D. S. acknowledge PhD studentship support from the BBSRC White Rose Mechanistic Biology DTP. This PhD was supported with generous industrial CASE support from Qkine and our thanks go to Dr Marko Hyv{\"o}nen and Dr Catherine Elton for their support. N. D. J. Y. and M. A. F. acknowledge support from the Engineering and Physical Sciences Research Council (EPSRC, EP/V044303/1). Dr Alex Heyam is thanked for help running NMR kinetics experiments. We thank Dr Ed Bergstrom and The York Centre of Excellence in Mass Spectrometry for support with protein mass spectrometry. This centre was created thanks to a major capital investment through Science City York, supported by Yorkshire Forward with funds from the Northern Way Initiative, and subsequent support from the EPSRC (EP/K039660/1; EP/M028127/1). Publisher Copyright: {\textcopyright} 2022 The Author(s).",

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AU - Barber, Lydia J.

AU - Yates, Nicholas D.J.

AU - Fascione, Martin A.

AU - Parkin, Alison

AU - Hemsworth, Glyn R.

AU - Genever, Paul G.

AU - Spicer, Christopher D.

N1 - Funding Information: L. J. B. and C. D. S. acknowledge PhD studentship support from the BBSRC White Rose Mechanistic Biology DTP. This PhD was supported with generous industrial CASE support from Qkine and our thanks go to Dr Marko Hyvönen and Dr Catherine Elton for their support. N. D. J. Y. and M. A. F. acknowledge support from the Engineering and Physical Sciences Research Council (EPSRC, EP/V044303/1). Dr Alex Heyam is thanked for help running NMR kinetics experiments. We thank Dr Ed Bergstrom and The York Centre of Excellence in Mass Spectrometry for support with protein mass spectrometry. This centre was created thanks to a major capital investment through Science City York, supported by Yorkshire Forward with funds from the Northern Way Initiative, and subsequent support from the EPSRC (EP/K039660/1; EP/M028127/1). Publisher Copyright: © 2022 The Author(s).

PY - 2022/11/17

Y1 - 2022/11/17

N2 - Protein N-termini provide uniquely reactive motifs for single site protein modification. Though a number of reactions have been developed to target this site, the selectivity, generality, and stability of the conjugates formed has not been studied. We have therefore undertaken a comprehensive comparative study of the most promising methods for N-terminal protein modification, and find that there is no ‘one size fits all’ approach, necessitating reagent screening for a particular protein or application. Moreover, we observed limited stability in all cases, leading to a need for continued innovation and development in the bioconjugation field.

AB - Protein N-termini provide uniquely reactive motifs for single site protein modification. Though a number of reactions have been developed to target this site, the selectivity, generality, and stability of the conjugates formed has not been studied. We have therefore undertaken a comprehensive comparative study of the most promising methods for N-terminal protein modification, and find that there is no ‘one size fits all’ approach, necessitating reagent screening for a particular protein or application. Moreover, we observed limited stability in all cases, leading to a need for continued innovation and development in the bioconjugation field.

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JO - RSC Chemical Biology

JF - RSC Chemical Biology

ER -

Selectivity and stability of N-terminal targeting protein modification chemistries

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Selectivity and stability of N-terminal targeting protein modification chemistries

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