Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring

Trine Amalie Fogh Gadeberg; Martin Høgholm Jørgensen; Heidi Gytz Olesen; Josefine Lorentzen; Seandean Lykke Harwood; Ana Viana Almeida; Marlene Uglebjerg Fruergaard; Rasmus Kjeldsen Jensen; Philipp Kanis; Henrik Pedersen; Emil Tranchant; Steen Vang Petersen; Ida Buch Thøgersen; Birthe Brandt Kragelund; Joseph Anthony Lyons; Jan Johannes Enghild; Gregers Rom Andersen

doi:10.1038/s41594-024-01467-4

Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring

Trine Amalie Fogh Gadeberg, Martin Høgholm Jørgensen, Heidi Gytz Olesen, Josefine Lorentzen, Seandean Lykke Harwood, Ana Viana Almeida, Marlene Uglebjerg Fruergaard, Rasmus Kjeldsen Jensen, Philipp Kanis, Henrik Pedersen, Emil Tranchant, Steen Vang Petersen, Ida Buch Thøgersen, Birthe Brandt Kragelund, Joseph Anthony Lyons, Jan Johannes Enghild, Gregers Rom Andersen^*

^*Corresponding author af dette arbejde

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

1 Citationer (Scopus)

Abstract

The C3 protein is the central molecule within the complement system and undergoes proteolytic activation to C3b in the presence of pathogens. Pattern-independent activation of C3 also occurs via hydrolysis, resulting in C3(H₂O), but the structural details of C3 hydrolysis remain elusive. Here we show that the conformation of the C3(H₂O) analog, C3MA, is indistinguishable from C3b. In contrast, the reaction intermediate C3* adopts a conformation dramatically different from both C3 and C3MA. In C3*, unlocking of the macroglobulin (MG) 3 domain creates a large opening in the MG ring through which the anaphylatoxin (ANA) domain translocates through a transient opening. C3MA formation is inhibited by an MG3-specific nanobody and prevented by linking the ANA domain to the C3 β-chain. Our study reveals an unexpected dynamic behavior of C3 and forms the basis for elucidation of the in vivo contribution of C3 hydrolysis and for controlling complement upon intravascular hemolysis and surface-contact-induced activation.

Originalsprog	Engelsk
Artikelnummer	872536
Tidsskrift	Nature Structural and Molecular Biology
Vol/bind	32
Nummer	5
Sider (fra-til)	884-895
Antal sider	12
ISSN	1545-9993
DOI	https://doi.org/10.1038/s41594-024-01467-4
Status	Udgivet - maj 2025

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Gadeberg, T. A. F., Jørgensen, M. H., Olesen, H. G., Lorentzen, J., Harwood, S. L., Almeida, A. V., Fruergaard, M. U., Jensen, R. K., Kanis, P., Pedersen, H., Tranchant, E., Petersen, S. V., Thøgersen, I. B., Kragelund, B. B., Lyons, J. A., Enghild, J. J., & Andersen, G. R. (2025). Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring. Nature Structural and Molecular Biology, 32(5), 884-895. Artikel 872536. https://doi.org/10.1038/s41594-024-01467-4

@article{6dcc8d7773aa43afa05fc83dafee4568,

title = "Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring",

abstract = "The C3 protein is the central molecule within the complement system and undergoes proteolytic activation to C3b in the presence of pathogens. Pattern-independent activation of C3 also occurs via hydrolysis, resulting in C3(H2O), but the structural details of C3 hydrolysis remain elusive. Here we show that the conformation of the C3(H2O) analog, C3MA, is indistinguishable from C3b. In contrast, the reaction intermediate C3* adopts a conformation dramatically different from both C3 and C3MA. In C3*, unlocking of the macroglobulin (MG) 3 domain creates a large opening in the MG ring through which the anaphylatoxin (ANA) domain translocates through a transient opening. C3MA formation is inhibited by an MG3-specific nanobody and prevented by linking the ANA domain to the C3 β-chain. Our study reveals an unexpected dynamic behavior of C3 and forms the basis for elucidation of the in vivo contribution of C3 hydrolysis and for controlling complement upon intravascular hemolysis and surface-contact-induced activation.",

author = "Gadeberg, \{Trine Amalie Fogh\} and J{\o}rgensen, \{Martin H{\o}gholm\} and Olesen, \{Heidi Gytz\} and Josefine Lorentzen and Harwood, \{Seandean Lykke\} and Almeida, \{Ana Viana\} and Fruergaard, \{Marlene Uglebjerg\} and Jensen, \{Rasmus Kjeldsen\} and Philipp Kanis and Henrik Pedersen and Emil Tranchant and Petersen, \{Steen Vang\} and Th{\o}gersen, \{Ida Buch\} and Kragelund, \{Birthe Brandt\} and Lyons, \{Joseph Anthony\} and Enghild, \{Jan Johannes\} and Andersen, \{Gregers Rom\}",

year = "2025",

month = may,

doi = "10.1038/s41594-024-01467-4",

language = "English",

volume = "32",

pages = "884--895",

journal = "Nature Structural and Molecular Biology",

issn = "1545-9993",

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number = "5",

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Gadeberg, TAF , Jørgensen, MH , Olesen, HG , Lorentzen, J , Harwood, SL, Almeida, AV, Fruergaard, MU, Jensen, RK, Kanis, P, Pedersen, H, Tranchant, E, Petersen, SV, Thøgersen, IB, Kragelund, BB, Lyons, JA , Enghild, JJ & Andersen, GR 2025, 'Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring', Nature Structural and Molecular Biology, bind 32, nr. 5, 872536, s. 884-895. https://doi.org/10.1038/s41594-024-01467-4

Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring. / Gadeberg, Trine Amalie Fogh ; Jørgensen, Martin Høgholm ; Olesen, Heidi Gytz et al.
I: Nature Structural and Molecular Biology, Bind 32, Nr. 5, 872536, 05.2025, s. 884-895.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avis › Tidsskriftartikel › Forskning › peer review

TY - JOUR

T1 - Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring

AU - Gadeberg, Trine Amalie Fogh

AU - Jørgensen, Martin Høgholm

AU - Olesen, Heidi Gytz

AU - Lorentzen, Josefine

AU - Harwood, Seandean Lykke

AU - Almeida, Ana Viana

AU - Fruergaard, Marlene Uglebjerg

AU - Jensen, Rasmus Kjeldsen

AU - Kanis, Philipp

AU - Pedersen, Henrik

AU - Tranchant, Emil

AU - Petersen, Steen Vang

AU - Thøgersen, Ida Buch

AU - Kragelund, Birthe Brandt

AU - Lyons, Joseph Anthony

AU - Enghild, Jan Johannes

AU - Andersen, Gregers Rom

PY - 2025/5

Y1 - 2025/5

N2 - The C3 protein is the central molecule within the complement system and undergoes proteolytic activation to C3b in the presence of pathogens. Pattern-independent activation of C3 also occurs via hydrolysis, resulting in C3(H2O), but the structural details of C3 hydrolysis remain elusive. Here we show that the conformation of the C3(H2O) analog, C3MA, is indistinguishable from C3b. In contrast, the reaction intermediate C3* adopts a conformation dramatically different from both C3 and C3MA. In C3*, unlocking of the macroglobulin (MG) 3 domain creates a large opening in the MG ring through which the anaphylatoxin (ANA) domain translocates through a transient opening. C3MA formation is inhibited by an MG3-specific nanobody and prevented by linking the ANA domain to the C3 β-chain. Our study reveals an unexpected dynamic behavior of C3 and forms the basis for elucidation of the in vivo contribution of C3 hydrolysis and for controlling complement upon intravascular hemolysis and surface-contact-induced activation.

AB - The C3 protein is the central molecule within the complement system and undergoes proteolytic activation to C3b in the presence of pathogens. Pattern-independent activation of C3 also occurs via hydrolysis, resulting in C3(H2O), but the structural details of C3 hydrolysis remain elusive. Here we show that the conformation of the C3(H2O) analog, C3MA, is indistinguishable from C3b. In contrast, the reaction intermediate C3* adopts a conformation dramatically different from both C3 and C3MA. In C3*, unlocking of the macroglobulin (MG) 3 domain creates a large opening in the MG ring through which the anaphylatoxin (ANA) domain translocates through a transient opening. C3MA formation is inhibited by an MG3-specific nanobody and prevented by linking the ANA domain to the C3 β-chain. Our study reveals an unexpected dynamic behavior of C3 and forms the basis for elucidation of the in vivo contribution of C3 hydrolysis and for controlling complement upon intravascular hemolysis and surface-contact-induced activation.

UR - http://www.scopus.com/inward/record.url?scp=85217207379&partnerID=8YFLogxK

U2 - 10.1038/s41594-024-01467-4

DO - 10.1038/s41594-024-01467-4

M3 - Journal article

C2 - 39849196

AN - SCOPUS:85217207379

SN - 1545-9993

VL - 32

SP - 884

EP - 895

JO - Nature Structural and Molecular Biology

JF - Nature Structural and Molecular Biology

IS - 5

M1 - 872536

ER -

Cryo-EM analysis of complement C3 reveals a reversible major opening of the macroglobulin ring

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