In vivo Cross-Linking MS of the Complement System MAC Assembled on Live Gram-Positive Bacteria - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Frontiers in Genetics Année : 2021

In vivo Cross-Linking MS of the Complement System MAC Assembled on Live Gram-Positive Bacteria

Résumé

Protein–protein interactions are central in many biological processes, but they are challenging to characterize, especially in complex samples. Protein cross-linking combined with mass spectrometry (MS) and computational modeling is gaining increased recognition as a viable tool in protein interaction studies. Here, we provide insights into the structure of the multicomponent human complement system membrane attack complex (MAC) using in vivo cross-linking MS combined with computational macromolecular modeling. We developed an affinity procedure followed by chemical cross-linking on human blood plasma using live Streptococcus pyogenes to enrich for native MAC associated with the bacterial surface. In this highly complex sample, we identified over 100 cross-linked lysine–lysine pairs between different MAC components that enabled us to present a quaternary model of the assembled MAC in its native environment. Demonstrating the validity of our approach, this MAC model is supported by existing X-ray crystallographic and electron cryo-microscopic models. This approach allows the study of protein–protein interactions in native environment mimicking their natural milieu. Its high potential in assisting and refining data interpretation in electron cryo-tomographic experiments will be discussed.
Fichier principal
Vignette du fichier
fgene-11-612475.pdf (2.16 Mo) Télécharger le fichier
Origine Fichiers éditeurs autorisés sur une archive ouverte

Dates et versions

hal-04429714 , version 1 (31-01-2024)

Identifiants

Citer

Hamed Khakzad, Lotta Happonen, Guy Tran van Nhieu, Johan Malmström, Lars Malmström. In vivo Cross-Linking MS of the Complement System MAC Assembled on Live Gram-Positive Bacteria. Frontiers in Genetics, 2021, 11, ⟨10.3389/fgene.2020.612475⟩. ⟨hal-04429714⟩
2 Consultations
3 Téléchargements

Altmetric

Partager

Gmail Mastodon Facebook X LinkedIn More