A C3-specific nanobody that blocks all three activation pathways in the human and murine complement system.
C3 convertase
antibody
complement system
inhibitor
innate immunity
nanobody
single-domain antibody
single-domain antibody (sdAb nanobody)
structural biology
Journal
The Journal of biological chemistry
ISSN: 1083-351X
Titre abrégé: J Biol Chem
Pays: United States
ID NLM: 2985121R
Informations de publication
Date de publication:
26 06 2020
26 06 2020
Historique:
received:
17
12
2019
revised:
28
04
2020
pubmed:
8
5
2020
medline:
14
1
2021
entrez:
8
5
2020
Statut:
ppublish
Résumé
The complement system is a tightly controlled proteolytic cascade in the innate immune system, which tags intruding pathogens and dying host cells for clearance. An essential protein in this process is complement component C3. Uncontrolled complement activation has been implicated in several human diseases and disorders and has spurred the development of therapeutic approaches that modulate the complement system. Here, using purified proteins and several biochemical assays and surface plasmon resonance, we report that our nanobody, hC3Nb2, inhibits C3 deposition by all complement pathways. We observe that the hC3Nb2 nanobody binds human native C3 and its degradation products with low nanomolar affinity and does not interfere with the endogenous regulation of C3b deposition mediated by Factors H and I. Using negative stain EM analysis and functional assays, we demonstrate that hC3Nb2 inhibits the substrate-convertase interaction by binding to the MG3 and MG4 domains of C3 and C3b. Furthermore, we notice that hC3Nb2 is cross-reactive and inhibits the lectin and alternative pathway in murine serum. We conclude that hC3Nb2 is a potent, general, and versatile inhibitor of the human and murine complement cascades. Its cross-reactivity suggests that this nanobody may be valuable for analysis of complement activation within animal models of both acute and chronic diseases.
Identifiants
pubmed: 32376685
pii: S0021-9258(17)49370-7
doi: 10.1074/jbc.RA119.012339
pmc: PMC7324514
pii:
doi:
Substances chimiques
Complement C3
0
Single-Domain Antibodies
0
Complement C3-C5 Convertases
EC 3.4.21.-
Banques de données
PDB
['2A74', '6EHG', '2XWJ', '5O32', '2A73', '2B39', '5FO7', '6RU5']
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
8746-8758Informations de copyright
© 2020 Pedersen et al.
Déclaration de conflit d'intérêts
Conflict of interest—G. R. A. has a collaboration with Alexion pharmaceuticals.
Références
Hum Genomics. 2015 Jun 10;9:7
pubmed: 26054645
Blood. 2009 Sep 24;114(13):2837-45
pubmed: 19584399
Front Immunol. 2019 Aug 22;10:2007
pubmed: 31507604
Proteins. 2013 Nov;81(11):1857-61
pubmed: 23852738
Nat Biotechnol. 2007 Nov;25(11):1256-64
pubmed: 17989688
Proc Natl Acad Sci U S A. 2012 Mar 20;109(12):4586-91
pubmed: 22393018
Mol Immunol. 2010 Aug;47(13):2154-60
pubmed: 20542571
Clin Exp Immunol. 2004 Dec;138(3):439-46
pubmed: 15544620
Immunobiology. 2016 Jun;221(6):733-9
pubmed: 26742632
Immunol Lett. 2017 Sep;189:64-72
pubmed: 28554712
EMBO J. 2011 Feb 2;30(3):606-16
pubmed: 21217642
Front Immunol. 2019 Jun 14;10:1157
pubmed: 31258525
Front Immunol. 2015 May 26;6:257
pubmed: 26074922
J Struct Biol. 2009 Apr;166(1):95-102
pubmed: 19263523
J Struct Biol. 1999 Dec 1;128(1):82-97
pubmed: 10600563
Cell Host Microbe. 2016 Jul 13;20(1):36-48
pubmed: 27345696
Eur J Clin Invest. 2015 Apr;45(4):423-40
pubmed: 25678219
Elife. 2018 Mar 07;7:
pubmed: 29513216
Mol Immunol. 2008 Apr;45(8):2370-9
pubmed: 18096230
Mol Immunol. 2019 Oct;114:341-352
pubmed: 31446305
Mol Immunol. 2013 Dec 15;56(3):222-31
pubmed: 23810291
J Struct Biol. 2009 May;166(2):205-13
pubmed: 19374019
Science. 2016 May 6;352(6286):712-716
pubmed: 27033548
Annu Rev Neurosci. 2012;35:369-89
pubmed: 22715882
Nature. 2005 Sep 22;437(7058):505-11
pubmed: 16177781
EMBO J. 2017 Apr 13;36(8):1084-1099
pubmed: 28264884
Proc Natl Acad Sci U S A. 2010 Feb 23;107(8):3681-6
pubmed: 20133685
Nature. 2006 Nov 9;444(7116):217-20
pubmed: 17051150
Mol Immunol. 2016 Aug;76:13-21
pubmed: 27337595
Mol Immunol. 2009 Dec;47(2-3):373-80
pubmed: 19800125
Cell. 2006 Mar 10;124(5):915-27
pubmed: 16530040
Nat Struct Mol Biol. 2017 Aug;24(8):643-651
pubmed: 28671664
Nature. 2016 Feb 11;530(7589):177-83
pubmed: 26814963
Proc Natl Acad Sci U S A. 2013 Oct 8;110(41):16426-31
pubmed: 24065820
J Biol Chem. 2009 Jan 30;284(5):3273-84
pubmed: 19010777
Nat Rev Drug Discov. 2019 Sep;18(9):707-729
pubmed: 31324874
Immunol Rev. 2001 Apr;180:112-22
pubmed: 11414353
J Hum Hypertens. 2007 Apr;21(4):276-82
pubmed: 17167524
Science. 2011 Apr 29;332(6029):608-11
pubmed: 21527715
Drug Des Devel Ther. 2019 Apr 17;13:1251-1258
pubmed: 31118566
Front Immunol. 2018 Nov 29;9:2822
pubmed: 30555486
Arthritis Rheum. 2006 Jun;54(6):1856-66
pubmed: 16736523
J Struct Biol. 2005 Jul;151(1):41-60
pubmed: 15890530
Science. 2010 Dec 24;330(6012):1816-20
pubmed: 21205667
J Biol Chem. 2007 Oct 5;282(40):29241-7
pubmed: 17684013
J Mol Biol. 2006 Aug 4;361(1):115-27
pubmed: 16831446
Semin Thromb Hemost. 2014 Jun;40(4):422-30
pubmed: 24799305
Nat Rev Nephrol. 2019 Mar;15(3):129-143
pubmed: 30692664
Nat Rev Dis Primers. 2017 May 18;3:17028
pubmed: 28516949
J Biol Chem. 2018 Apr 27;293(17):6269-6281
pubmed: 29497000
EMBO J. 2015 Nov 12;34(22):2735-57
pubmed: 26489954
EMBO J. 2016 May 17;35(10):1133-49
pubmed: 27013439
Immunol Rev. 2016 Nov;274(1):59-73
pubmed: 27782336
Immunity. 2012 Aug 24;37(2):199-207
pubmed: 22921118
J Innate Immun. 2018;10(5-6):455-464
pubmed: 30149378
J Struct Biol. 2012 Dec;180(3):519-30
pubmed: 23000701