Native and Ion Mobility Mass Spectrometry Characterization of Alpha 1 Antitrypsin Variants and Oligomers

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2023 Dec 18

PMID 38108966

Authors

Sarah Vickers

James Irving

David A Lomas

Konstantinos Thalassinos

Affiliations

Soon will be listed here.

Abstract

In this chapter, we describe a method for analyzing both recombinant and plasma-derived alpha 1 antitrypsin and its oligomers by means of native ion mobility mass spectrometry. Our experimental workflow can be applied to other variants of alpha 1 antitrypsin and its oligomers as well as being used to probe their interactions with small molecules in the gas phase.

References

Gabelica V, Shvartsburg A, Afonso C, Barran P, Benesch J, Bleiholder C . Recommendations for reporting ion mobility Mass Spectrometry measurements. Mass Spectrom Rev. 2019; 38(3):291-320. PMC: 6618043. DOI: 10.1002/mas.21585. View

Hopper J, Oldham N . Collision induced unfolding of protein ions in the gas phase studied by ion mobility-mass spectrometry: the effect of ligand binding on conformational stability. J Am Soc Mass Spectrom. 2009; 20(10):1851-8. DOI: 10.1016/j.jasms.2009.06.010. View

Powell L, Pain R . Effects of glycosylation on the folding and stability of human, recombinant and cleaved alpha 1-antitrypsin. J Mol Biol. 1992; 224(1):241-52. DOI: 10.1016/0022-2836(92)90587-a. View

Lomas D, Irving J, Arico-Muendel C, Belyanskaya S, Brewster A, Brown M . Development of a small molecule that corrects misfolding and increases secretion of Z α -antitrypsin. EMBO Mol Med. 2021; 13(3):e13167. PMC: 7933930. DOI: 10.15252/emmm.202013167. View

Carrell R, Lomas D . Alpha1-antitrypsin deficiency--a model for conformational diseases. N Engl J Med. 2002; 346(1):45-53. DOI: 10.1056/NEJMra010772. View

Faull S, Elliston E, Gooptu B, Jagger A, Aldobiyan I, Redzej A . The structural basis for Z α-antitrypsin polymerization in the liver. Sci Adv. 2020; 6(43). PMC: 7577719. DOI: 10.1126/sciadv.abc1370. View

Nyon M, Prentice T, Day J, Kirkpatrick J, Sivalingam G, Levy G . An integrative approach combining ion mobility mass spectrometry, X-ray crystallography, and nuclear magnetic resonance spectroscopy to study the conformational dynamics of α1 -antitrypsin upon ligand binding. Protein Sci. 2015; 24(8):1301-12. PMC: 4534181. DOI: 10.1002/pro.2706. View

Giles K, Ujma J, Wildgoose J, Pringle S, Richardson K, Langridge D . A Cyclic Ion Mobility-Mass Spectrometry System. Anal Chem. 2019; 91(13):8564-8573. DOI: 10.1021/acs.analchem.9b01838. View

10.

Eldrid C, Ben-Younis A, Ujma J, Britt H, Cragnolini T, Kalfas S . Cyclic Ion Mobility-Collision Activation Experiments Elucidate Protein Behavior in the Gas Phase. J Am Soc Mass Spectrom. 2021; 32(6):1545-1552. PMC: 8172447. DOI: 10.1021/jasms.1c00018. View

11.

Jagger A, Waudby C, Irving J, Christodoulou J, Lomas D . High-resolution ex vivo NMR spectroscopy of human Z α-antitrypsin. Nat Commun. 2020; 11(1):6371. PMC: 7732992. DOI: 10.1038/s41467-020-20147-7. View

12.

Haq I, Irving J, Saleh A, Dron L, Regan-Mochrie G, Motamedi-Shad N . Deficiency Mutations of Alpha-1 Antitrypsin. Effects on Folding, Function, and Polymerization. Am J Respir Cell Mol Biol. 2015; 54(1):71-80. PMC: 4742932. DOI: 10.1165/rcmb.2015-0154OC. View

13.

Chang Y, Mahadeva R, Chang W, Lin S, Chu Y . Small-molecule peptides inhibit Z alpha1-antitrypsin polymerization. J Cell Mol Med. 2009; 13(8B):2304-2316. PMC: 6529975. DOI: 10.1111/j.1582-4934.2008.00608.x. View

14.

Eldrid C, Ujma J, Kalfas S, Tomczyk N, Giles K, Morris M . Gas Phase Stability of Protein Ions in a Cyclic Ion Mobility Spectrometry Traveling Wave Device. Anal Chem. 2019; 91(12):7554-7561. PMC: 7006968. DOI: 10.1021/acs.analchem.8b05641. View

15.

Kanu A, Dwivedi P, Tam M, Matz L, Hill Jr H . Ion mobility-mass spectrometry. J Mass Spectrom. 2008; 43(1):1-22. DOI: 10.1002/jms.1383. View

16.

Polasky D, Dixit S, Fantin S, Ruotolo B . CIUSuite 2: Next-Generation Software for the Analysis of Gas-Phase Protein Unfolding Data. Anal Chem. 2019; 91(4):3147-3155. DOI: 10.1021/acs.analchem.8b05762. View