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 journalArticlepeer-review

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 languageEnglish
Article number63586
JournaleLife
DOIs
Publication statusPublished - 11 Feb 2021

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