Determination of hATG8 Binding Selectivity of AIM (Autophagy-Interacting Motif) Peptides Using Native Electrospray Ionization Mass Spectrometry

Andrew Brennan; Neil Oldham; Robert Layfield; Daniel Scott

doi:10.1007/978-1-0716-4067-8_19

Determination of hATG8 Binding Selectivity of AIM (Autophagy-Interacting Motif) Peptides Using Native Electrospray Ionization Mass Spectrometry

Andrew Brennan, Neil Oldham, Robert Layfield, Daniel Scott

Department of Life Sciences

University of Nottingham

Research output: Chapter or section in a book/report/conference proceeding › Chapter or section

Abstract

Establishing the hATG8 binding selectivity of AIM (autophagy-interacting motif) sequences found within autophagy system proteins provides insights into their biological roles, and in the case of disease-associated AIM mutations, potential pathophysiological mechanisms. Given the sometimes small differences in affinity for an individual AIM amongst the six hATG8 proteins, establishing AIM preferences can be experimentally challenging. We describe a native mass spectrometry method that is suitable for detecting such differences, using synthetic AIM peptides and recombinant hATG8 proteins, to probe hATG8-AIM interactions in the gas phase. Binding preferences of a single AIM peptide against multiple hATG8s, or two AIM peptides against a single hATG8 (e.g., wild-type versus mutant AIM), may be determined.

Original language	English
Title of host publication	Selective Autophagy. Methods in Molecular Biology
Editors	I. P. Nezis
Place of Publication	New York, U. S. A.
Publisher	Humana Press
Pages	237-246
Number of pages	10
ISBN (Electronic)	9781071640678
ISBN (Print)	9781071640661
DOIs	https://doi.org/10.1007/978-1-0716-4067-8_19
Publication status	Published - 9 Aug 2024

Publication series

Name	Methods in Molecular Biology
Volume	2845

Keywords

Autophagy-interacting motif
Binding selectivity
Native mass spectrometry
hATG8

ASJC Scopus subject areas

Genetics
Molecular Biology

Access to Document

10.1007/978-1-0716-4067-8_19

Cite this

Brennan, A., Oldham, N., Layfield, R., & Scott, D. (2024). Determination of hATG8 Binding Selectivity of AIM (Autophagy-Interacting Motif) Peptides Using Native Electrospray Ionization Mass Spectrometry. In I. P. Nezis (Ed.), Selective Autophagy. Methods in Molecular Biology (pp. 237-246). (Methods in Molecular Biology ; Vol. 2845). Humana Press. https://doi.org/10.1007/978-1-0716-4067-8_19

Determination of hATG8 Binding Selectivity of AIM (Autophagy-Interacting Motif) Peptides Using Native Electrospray Ionization Mass Spectrometry. / Brennan, Andrew; Oldham, Neil; Layfield, Robert et al.
Selective Autophagy. Methods in Molecular Biology. ed. / I. P. Nezis. New York, U. S. A.: Humana Press, 2024. p. 237-246 (Methods in Molecular Biology ; Vol. 2845).

Research output: Chapter or section in a book/report/conference proceeding › Chapter or section

Brennan, A, Oldham, N, Layfield, R & Scott, D 2024, Determination of hATG8 Binding Selectivity of AIM (Autophagy-Interacting Motif) Peptides Using Native Electrospray Ionization Mass Spectrometry. in IP Nezis (ed.), Selective Autophagy. Methods in Molecular Biology. Methods in Molecular Biology , vol. 2845, Humana Press, New York, U. S. A., pp. 237-246. https://doi.org/10.1007/978-1-0716-4067-8_19

Brennan A, Oldham N, Layfield R, Scott D. Determination of hATG8 Binding Selectivity of AIM (Autophagy-Interacting Motif) Peptides Using Native Electrospray Ionization Mass Spectrometry. In Nezis IP, editor, Selective Autophagy. Methods in Molecular Biology. New York, U. S. A.: Humana Press. 2024. p. 237-246. (Methods in Molecular Biology ). doi: 10.1007/978-1-0716-4067-8_19

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Determination of hATG8 Binding Selectivity of AIM (Autophagy-Interacting Motif) Peptides Using Native Electrospray Ionization Mass Spectrometry

Abstract

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