In vivo Cross-Linking MS of the Complement System MAC Assembled on Live Gram-Positive Bacteria
Khakzad, Hamed ; Happonen, Lotta LU ; Tran Van Nhieu, Guy ; Malmström, Johan LU
and Malmström, Lars
LU
(2021)
In Frontiers in Genetics
11.
- Abstract
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... (More)
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.
(Less)
- author
- Khakzad, Hamed
; Happonen, Lotta
LU
; Tran Van Nhieu, Guy
; Malmström, Johan
LU
and Malmström, Lars
LU
- organization
- publishing date
- 2021-01-08
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Frontiers in Genetics
- volume
- 11
- article number
- 612475
- publisher
- Frontiers Media S. A.
- external identifiers
-
- pmid:33488677
- scopus:85099761856
- ISSN
- 1664-8021
- DOI
- 10.3389/fgene.2020.612475
- language
- English
- LU publication?
- yes
- additional info
- Copyright © 2021 Khakzad, Happonen, Tran Van Nhieu, Malmström and Malmström.
- id
- 2ac8cfc7-be45-43f1-8340-c20d9a027512
- date added to LUP
- 2021-01-31 23:14:34
- date last changed
- 2024-06-27 07:50:07
@article{2ac8cfc7-be45-43f1-8340-c20d9a027512,
abstract = {{<p>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.</p>}},
author = {{Khakzad, Hamed and Happonen, Lotta and Tran Van Nhieu, Guy and Malmström, Johan and Malmström, Lars}},
issn = {{1664-8021}},
language = {{eng}},
month = {{01}},
publisher = {{Frontiers Media S. A.}},
series = {{Frontiers in Genetics}},
title = {{In vivo Cross-Linking MS of the Complement System MAC Assembled on Live Gram-Positive Bacteria}},
url = {{http://dx.doi.org/10.3389/fgene.2020.612475}},
doi = {{10.3389/fgene.2020.612475}},
volume = {{11}},
year = {{2021}},
}