The allosteric modulation of Complement C5 by knob domain peptides

Alex Macpherson; Maisem Laabei; Zainab Ahdash; Melissa Graewert; James Birtley; Monika-Sarah Schulze; Susan Crennell; Sarah Robinson; Ben Holmes; Vladas Oleinikovas; Per Nilsson; James Snowdon; Victoria Ellis; Tom Eirik Mollnes; Charlotte Deane; Dmitri Svergun; Alastair Lawson; Jean Van Den Elsen

doi:10.7554/eLife.63586

The allosteric modulation of Complement C5 by knob domain peptides

Alex Macpherson, Maisem Laabei, Zainab Ahdash, Melissa Graewert, James Birtley, Monika-Sarah Schulze, Susan Crennell, Sarah Robinson, Ben Holmes, Vladas Oleinikovas, Per Nilsson, James Snowdon, Victoria Ellis, Tom Eirik Mollnes, Charlotte Deane, Dmitri Svergun, Alastair Lawson, Jean Van Den Elsen

Research output: Contribution to journal › Article › peer-review

21 Citations (SciVal)

Abstract

Bovines have evolved a subset of antibodies with ultra-long CDRH3 regions that harbour cysteine-rich knob domains. To produce high affinity peptides, we previously isolated autonomous 3-6 kDa knob domains from bovine antibodies. Here, we show that binding of four knob domain peptides elicits a range of effects on the clinically validated drug target complement C5. Allosteric mechanisms predominated, with one peptide selectively inhibiting C5 cleavage by the alternative pathway C5 convertase, revealing a targetable mechanistic difference between the classical and alternative pathway C5 convertases. Taking a hybrid biophysical approach, we present C5-knob domain cocrystal structures and, by solution methods, observed allosteric effects propagating >50 Å from the binding sites. This study expands the therapeutic scope of C5, presents new inhibitors and introduces knob domains as new, low molecular weight antibody fragments, with therapeutic potential.

Original language	English
Article number	e63586
Pages (from-to)	1-49
Number of pages	49
Journal	eLife
Volume	10
DOIs	https://doi.org/10.7554/eLife.63586
Publication status	Published - 11 Feb 2021

Bibliographical note

Funding Information:
This work was performed by A.M. as partial fulfilment of the requirements for a PhD from the University of Bath and was funded by UCB. A.M., S.S., V.O., J.S., B.H., V.E., Z.A. and A.D.G.L are present or past employees of UCB and may hold shares and/or stock options. T.E.M is a Board member of Ra Pharmaceuticals, Inc. All other authors declare no competing interests.

Data availability
Structural datasets presented in this study have been made publicly available in the Protein Data
Bank (PDB) and Small Angle Scattering Biological Data Bank (SASBDB).
The following datasets were generated:
Author(s) Year Dataset title Dataset URL
Database and
Identifier
Macpherson A,
Elsen JM
2021 Crystal structure C5-K8 complex https://www.rcsb.org/
structure/7AD7
RCSB Protein Data
Bank, 7AD7
Macpherson A,
Elsen JM
2021 Crystal structure of C5-K92
complex
https://www.rcsb.org/
structure/7AD6
RCSB Protein Data
Bank, 7AD6
Macpherson A,
Elsen JM, Graewert
MA, Svergun D
2020 SAXS data and models of C5-
bovine knob domain peptides
https://www.sasbdb.org/
data/SASDJA6/
Small Angle
Scattering Biological
Data Bank, SASDJA6

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10.7554/eLife.63586Licence: CC BY

Cite this

Macpherson, A., Laabei, M., Ahdash, Z., Graewert, M., Birtley, J., Schulze, M.-S., Crennell, S., Robinson, S., Holmes, B., Oleinikovas, V., Nilsson, P., Snowdon, J., Ellis, V., Mollnes, T. E., Deane, C., Svergun, D., Lawson, A., & Van Den Elsen, J. (2021). The allosteric modulation of Complement C5 by knob domain peptides. eLife, 10, 1-49. Article e63586. https://doi.org/10.7554/eLife.63586

Macpherson, A, Laabei, M, Ahdash, Z, Graewert, M, Birtley, J, Schulze, M-S, Crennell, S, Robinson, S, Holmes, B, Oleinikovas, V, Nilsson, P, Snowdon, J, Ellis, V, Mollnes, TE, Deane, C, Svergun, D, Lawson, A & Van Den Elsen, J 2021, 'The allosteric modulation of Complement C5 by knob domain peptides', eLife, vol. 10, e63586, pp. 1-49. https://doi.org/10.7554/eLife.63586

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title = "The allosteric modulation of Complement C5 by knob domain peptides",

abstract = "Bovines have evolved a subset of antibodies with ultra-long CDRH3 regions that harbour cysteine-rich knob domains. To produce high affinity peptides, we previously isolated autonomous 3-6 kDa knob domains from bovine antibodies. Here, we show that binding of four knob domain peptides elicits a range of effects on the clinically validated drug target complement C5. Allosteric mechanisms predominated, with one peptide selectively inhibiting C5 cleavage by the alternative pathway C5 convertase, revealing a targetable mechanistic difference between the classical and alternative pathway C5 convertases. Taking a hybrid biophysical approach, we present C5-knob domain cocrystal structures and, by solution methods, observed allosteric effects propagating >50 {\AA} from the binding sites. This study expands the therapeutic scope of C5, presents new inhibitors and introduces knob domains as new, low molecular weight antibody fragments, with therapeutic potential.",

author = "Alex Macpherson and Maisem Laabei and Zainab Ahdash and Melissa Graewert and James Birtley and Monika-Sarah Schulze and Susan Crennell and Sarah Robinson and Ben Holmes and Vladas Oleinikovas and Per Nilsson and James Snowdon and Victoria Ellis and Mollnes, {Tom Eirik} and Charlotte Deane and Dmitri Svergun and Alastair Lawson and {Van Den Elsen}, Jean",

note = "Funding Information: This work was performed by A.M. as partial fulfilment of the requirements for a PhD from the University of Bath and was funded by UCB. A.M., S.S., V.O., J.S., B.H., V.E., Z.A. and A.D.G.L are present or past employees of UCB and may hold shares and/or stock options. T.E.M is a Board member of Ra Pharmaceuticals, Inc. All other authors declare no competing interests. Data availability Structural datasets presented in this study have been made publicly available in the Protein Data Bank (PDB) and Small Angle Scattering Biological Data Bank (SASBDB). The following datasets were generated: Author(s) Year Dataset title Dataset URL Database and Identifier Macpherson A, Elsen JM 2021 Crystal structure C5-K8 complex https://www.rcsb.org/ structure/7AD7 RCSB Protein Data Bank, 7AD7 Macpherson A, Elsen JM 2021 Crystal structure of C5-K92 complex https://www.rcsb.org/ structure/7AD6 RCSB Protein Data Bank, 7AD6 Macpherson A, Elsen JM, Graewert MA, Svergun D 2020 SAXS data and models of C5- bovine knob domain peptides https://www.sasbdb.org/ data/SASDJA6/ Small Angle Scattering Biological Data Bank, SASDJA6 ",

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AU - Macpherson, Alex

AU - Laabei, Maisem

AU - Ahdash, Zainab

AU - Graewert, Melissa

AU - Birtley, James

AU - Schulze, Monika-Sarah

AU - Crennell, Susan

AU - Robinson, Sarah

AU - Holmes, Ben

AU - Oleinikovas, Vladas

AU - Nilsson, Per

AU - Snowdon, James

AU - Ellis, Victoria

AU - Mollnes, Tom Eirik

AU - Deane, Charlotte

AU - Svergun, Dmitri

AU - Lawson, Alastair

AU - Van Den Elsen, Jean

N1 - Funding Information: This work was performed by A.M. as partial fulfilment of the requirements for a PhD from the University of Bath and was funded by UCB. A.M., S.S., V.O., J.S., B.H., V.E., Z.A. and A.D.G.L are present or past employees of UCB and may hold shares and/or stock options. T.E.M is a Board member of Ra Pharmaceuticals, Inc. All other authors declare no competing interests. Data availability Structural datasets presented in this study have been made publicly available in the Protein Data Bank (PDB) and Small Angle Scattering Biological Data Bank (SASBDB). The following datasets were generated: Author(s) Year Dataset title Dataset URL Database and Identifier Macpherson A, Elsen JM 2021 Crystal structure C5-K8 complex https://www.rcsb.org/ structure/7AD7 RCSB Protein Data Bank, 7AD7 Macpherson A, Elsen JM 2021 Crystal structure of C5-K92 complex https://www.rcsb.org/ structure/7AD6 RCSB Protein Data Bank, 7AD6 Macpherson A, Elsen JM, Graewert MA, Svergun D 2020 SAXS data and models of C5- bovine knob domain peptides https://www.sasbdb.org/ data/SASDJA6/ Small Angle Scattering Biological Data Bank, SASDJA6

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AB - Bovines have evolved a subset of antibodies with ultra-long CDRH3 regions that harbour cysteine-rich knob domains. To produce high affinity peptides, we previously isolated autonomous 3-6 kDa knob domains from bovine antibodies. Here, we show that binding of four knob domain peptides elicits a range of effects on the clinically validated drug target complement C5. Allosteric mechanisms predominated, with one peptide selectively inhibiting C5 cleavage by the alternative pathway C5 convertase, revealing a targetable mechanistic difference between the classical and alternative pathway C5 convertases. Taking a hybrid biophysical approach, we present C5-knob domain cocrystal structures and, by solution methods, observed allosteric effects propagating >50 Å from the binding sites. This study expands the therapeutic scope of C5, presents new inhibitors and introduces knob domains as new, low molecular weight antibody fragments, with therapeutic potential.

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The allosteric modulation of Complement C5 by knob domain peptides

Abstract

Bibliographical note

Access to Document

Other files and links

Fingerprint

Dissecting complement resistance in Staphylococcus aureus

HPLC-MS

Ultra High Pressure Liquid Chromatography instrument (UPLC) - used in conjunction with other DReaM instruments

Cite this

The allosteric modulation of Complement C5 by knob domain peptides

Abstract

Bibliographical note

Access to Document

Other files and links

Fingerprint

Projects

Dissecting complement resistance in Staphylococcus aureus

Equipment

HPLC-MS

Ultra High Pressure Liquid Chromatography instrument (UPLC) - used in conjunction with other DReaM instruments

Cite this