An inhibitor of complement C5 provides structural insights into activation

Significance The complement system is a crucial antimicrobial system in the human body. However, controlling its regulation is essential, and failure to do so is implicated in a number of clinical inflammatory pathologies leading to great interest in therapeutic complement inhibition. We have identified and characterized a class of complement inhibitors from biting ticks. Utilizing both cryoelectron microscopy and X-ray crystallography we provide a comprehensive understanding of their mechanism of inhibition at the level of the terminal pathway of complement. We present a high-resolution cryoelectron microscopy structure of complement C5, the molecule targeted by the major therapeutic Eculizumab. In addition, we reveal the fold of the CirpT family of tick inhibitors and their unique mode of inhibition.
The complement system is a crucial part of innate immune defenses against invading pathogens. The blood-meal of the tick Rhipicephalus pulchellus lasts for days, and the tick must therefore rely on inhibitors to counter complement activation. We have identified a class of inhibitors from tick saliva, the CirpT family, and generated detailed structural data revealing their mechanism of action. We show direct binding of a CirpT to complement C5 and have determined the structure of the C5–CirpT complex by cryoelectron microscopy. This reveals an interaction with the peripheral macro globulin domain 4 (C5_MG4) of C5. To achieve higher resolution detail, the structure of the C5_MG4–CirpT complex was solved by X-ray crystallography (at 2.7 Å). We thus present the fold of the CirpT protein family, and provide detailed mechanistic insights into its inhibitory function. Analysis of the binding interface reveals a mechanism of C5 inhibition, and provides information to expand our biological understanding of the activation of C5, and thus the terminal complement pathway.