Journal
FRONTIERS IN GENETICS
Volume 11, Issue -, Pages -Publisher
FRONTIERS MEDIA SA
DOI: 10.3389/fgene.2020.612475
Keywords
membrane attack complex; cross-linking mass spectrometry; Streptococcus pyogenes; Gram-positive bacteria; in vivo cross-linking
Categories
Funding
- Swiss National Science Foundation [P2ZHP3_191289]
- Foundation of Knut and Alice Wallenberg [2016.0023, 2019.0353]
- Swiss National Science Foundation (SNF) [P2ZHP3_191289] Funding Source: Swiss National Science Foundation (SNF)
Ask authors/readers for more resources
In this study, protein-protein interactions within the human complement system membrane attack complex (MAC) were investigated using in vivo cross-linking mass spectrometry and computational modeling. Over 100 cross-linked lysine-lysine pairs were identified between different MAC components, leading to the proposal of a quaternary model of the assembled MAC in its native environment. The approach demonstrated the validity of the MAC model through comparison with existing X-ray crystallographic and electron cryo-microscopic models, showcasing the potential of this method in studying native protein interactions.
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available