Native Ambient Mass Spectrometry of an Intact Membrane Protein Assembly and Soluble Protein Assemblies Directly from Lens Tissue.
Angew Chem Weinheim Bergstr Ger
; 134(31): e202201458, 2022 Aug 01.
Article
en En
| MEDLINE
| ID: mdl-38505128
ABSTRACT
Membrane proteins constitute around two-thirds of therapeutic targets but present a significant challenge for structural analysis due to their low abundance and solubility. Existing methods for structural analysis rely on over-expression and/or purification of the membrane protein, thus removing any links back to actual physiological environment. Here, we demonstrate mass spectrometry analysis of an intact oligomeric membrane protein directly from tissue. Aquaporin-0 exists as a 113â
kDa tetramer, with each subunit featuring six transmembrane helices. We report the characterisation of the intact assembly directly from a section of sheep eye lens without sample pre-treatment. Protein identity was confirmed by mass measurement of the tetramer and subunits, together with top-down mass spectrometry, and the spatial distribution was determined by mass spectrometry imaging. Our approach allows simultaneous analysis of soluble protein assemblies in the tissue.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Idioma:
En
Revista:
Angew Chem Weinheim Bergstr Ger
/
Angew. chem. (Weinheim Bergdtr. Germ.)
/
Angewandte chemie (Weinheim an der Bergstrasse, Germany)
Asunto de la revista:
BIOFISICA
/
QUIMICA
Año:
2022
Tipo del documento:
Article