Structural basis of complement membrane attack complex formation

Serna, Marina; Giles, Joanna L.; Morgan, B. Paul; Bubeck, Doryen

doi:10.1038/ncomms10587

Cited by 208 publications

(258 citation statements)

References 41 publications

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“…To determine whether serum complement components contribute to the observed killing of K. pneumoniae in serum, we first measured the surface association of C5b to C9 (C5b-C9), complement molecules that form the membrane attack complex (MAC) (33), by flow cytometry (Fig. 4).…”

Section: Resultsmentioning

confidence: 99%

Survival of Carbapenem-Resistant Klebsiella pneumoniae Sequence Type 258 in Human Blood

DeLeo

Kobayashi

Porter

et al. 2017

Antimicrob Agents Chemother

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Klebsiella pneumoniae is a prominent cause of nosocomial infections worldwide. Bloodstream infections caused by carbapenem-resistant K. pneumoniae, including the epidemic lineage known as multilocus sequence type 258 (ST258), are difficult to treat, and the rate of mortality from such infections is high. Thus, it is imperative that we gain a better understanding of host defense against this pathogen as a step toward developing novel therapies. Here we tested the hypothesis that the resistance of ST258 to bactericidal components of human blood, such as serum complement, is linked to virulence capacity in the context of bacteremia. There was significant variance in the survival of ST258 clinical isolates in heparinized human blood or normal human serum. The rate of survival of ST258 isolates in human blood was, in general, similar to that in normal human serum, suggesting a prominent role for complement (rather than leukocytes) in the healthy host defense against ST258 isolates and related organisms. Indeed, deposition of serum complement-the C5b to C9 (C5b-C9) membrane attack complex-onto the surface of ST258 isolates accompanied serum bactericidal activity. Human serum treated with pharmacological inhibitors of complement, depleted of antibody, or heated at 56°C for 30 min had significantly reduced or absent bactericidal activity. In contrast to heparinized blood from humans, that from BALB/c mice lacked bactericidal activity toward the ST258 isolates tested, but the virulence of these ST258 isolates in a mouse bacteremia model was inexplicably limited. Our data highlight the importance of the complement system in host defense against ST258 bacteremia, and we propose that there is the potential to enhance complement-mediated bactericidal activity using an antibody-based approach.

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Section: Resultsmentioning

confidence: 99%

Survival of Carbapenem-Resistant Klebsiella pneumoniae Sequence Type 258 in Human Blood

DeLeo

Kobayashi

Porter

et al. 2017

Antimicrob Agents Chemother

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“…More recently, the structures of a detergent-solubilised MAC pore (Serna et al, 2016) and a soluble C9 oligomer (Dudkina et al, 2016) have been solved with cryoelectron microscopy analyses to a resolution of 7-8 Å (Fig. 3) -the highest resolution so far achieved for MACPF assemblies.…”

Section: Sonnen Et Al 2014mentioning

confidence: 99%

“…For the CDC proteins perfringolysin O and streptolysin O, the prepore-to-pore transition can be controlled by altering an electrostatic interaction between two residues on the subunit interface (Lys336 and Glu183 in perfringolysin O) (Wade et al, (Leung et al, 2014), (D) a pleurotolysin pore (Lukoyanova et al, 2015), (E) a membrane-bound MAC pore (reproduced from Serna et al, 2016), and (F) a soluble C9 oligomer (reproduced from Dudkina et al, 2016). Cut-away side views show fitted atomic models.…”

Section: Cryoelectron Microscopy Reveals Intermediate Steps Of Pore Fmentioning

confidence: 99%

The membrane attack complex, perforin and cholesterol-dependent cytolysin superfamily of pore-forming proteins

Lukoyanova

Hoogenboom

Saibil

2016

Journal of Cell Science

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The membrane attack complex and perforin proteins (MACPFs) and bacterial cholesterol-dependent cytolysins (CDCs) are two branches of a large and diverse superfamily of pore-forming proteins that function in immunity and pathogenesis. During pore formation, soluble monomers assemble into large transmembrane pores through conformational transitions that involve extrusion and refolding of two α-helical regions into transmembrane β-hairpins. These transitions entail a dramatic refolding of the protein structure, and the resulting assemblies create large holes in cellular membranes, but they do not use any external source of energy. Structures of the membrane-bound assemblies are required to mechanistically understand and modulate these processes. In this Commentary, we discuss recent advances in the understanding of assembly mechanisms and molecular details of the conformational changes that occur during MACPF and CDC pore formation.

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“…The data collected at eBIC have, at the time of writing, generated ten research publications (Hospenthal et al, 2016;Serna et al, 2016;Joseph et al, 2016;Wilkinson et al, 2016;Iadanza et al, 2016;Ramsay et al, 2016;Fica et al, 2017;Swuec et al, 2017;Ilangovan et al, 2017;Boland et al, 2017). We have also received a number of personal communications from inhouse and external users reporting reconstructions at better than 4 Å resolution, with a few extending beyond 3 Å .…”

Section: Results From the First Year Of Ebicmentioning

confidence: 99%

Electron Bio-Imaging Centre (eBIC): the UK national research facility for biological electron microscopy

Clare

Siebert

Hecksel

et al. 2017

Acta Cryst Sect D Struct Biol

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The recent resolution revolution in cryo-EM has led to a massive increase in demand for both time on high-end cryo-electron microscopes and access to cryoelectron microscopy expertise. In anticipation of this demand, eBIC was set up at Diamond Light Source in collaboration with Birkbeck College London and the University of Oxford, and funded by the Wellcome Trust, the UK Medical Research Council (MRC) and the Biotechnology and Biological Sciences Research Council (BBSRC) to provide access to high-end equipment through peer review. eBIC is currently in its start-up phase and began by offering time on a single FEI Titan Krios microscope equipped with the latest generation of direct electron detectors from two manufacturers. Here, the current status and modes of access for potential users of eBIC are outlined. In the first year of operation, 222 d of microscope time were delivered to external research groups, with 95 visits in total, of which 53 were from unique groups. The data collected have generated multiple high-to intermediate-resolution structures (2.8-8 Å ), ten of which have been published. A second Krios microscope is now in operation, with two more due to come online in 2017. In the next phase of growth of eBIC, in addition to more microscope time, new data-collection strategies and sample-preparation techniques will be made available to external user groups. Finally, all raw data are archived, and a metadata catalogue and automated pipelines for data analysis are being developed.

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Structural basis of complement membrane attack complex formation

Cited by 208 publications

References 41 publications

Survival of Carbapenem-Resistant Klebsiella pneumoniae Sequence Type 258 in Human Blood

Survival of Carbapenem-Resistant Klebsiella pneumoniae Sequence Type 258 in Human Blood

The membrane attack complex, perforin and cholesterol-dependent cytolysin superfamily of pore-forming proteins

Electron Bio-Imaging Centre (eBIC): the UK national research facility for biological electron microscopy

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