Journal ArticleDOI
The structural basis for membrane binding and pore formation by lymphocyte perforin
Ruby H. P. Law,Natalya Lukoyanova,Ilia Voskoboinik,Ilia Voskoboinik,Tom T. Caradoc-Davies,Katherine Baran,Michelle A. Dunstone,Michelle A. Dunstone,Michael E. D'Angelo,Elena V. Orlova,Fasséli Coulibaly,Sandra Verschoor,Kylie A. Browne,Annette Ciccone,Michael J. Kuiper,Phillip I. Bird,Joseph A. Trapani,Joseph A. Trapani,Helen R. Saibil,James C. Whisstock,James C. Whisstock +20 more
TLDR
In this article, the X-ray crystal structure of perforin pore formation was determined using a cryo-electron microscopy reconstruction, which revealed remarkable flexibility in the mechanism of action and provided new insights into how related immune defence molecules such as complement proteins assemble into pores.Abstract:
Natural killer cells and cytotoxic T lymphocytes accomplish the critically important function of killing virus-infected and neoplastic cells. They do this by releasing the pore-forming protein perforin and granzyme proteases from cytoplasmic granules into the cleft formed between the abutting killer and target cell membranes. Perforin, a 67-kilodalton multidomain protein, oligomerizes to form pores that deliver the pro-apoptopic granzymes into the cytosol of the target cell. The importance of perforin is highlighted by the fatal consequences of congenital perforin deficiency, with more than 50 different perforin mutations linked to familial haemophagocytic lymphohistiocytosis (type 2 FHL). Here we elucidate the mechanism of perforin pore formation by determining the X-ray crystal structure of monomeric murine perforin, together with a cryo-electron microscopy reconstruction of the entire perforin pore. Perforin is a thin 'key-shaped' molecule, comprising an amino-terminal membrane attack complex perforin-like (MACPF)/cholesterol dependent cytolysin (CDC) domain followed by an epidermal growth factor (EGF) domain that, together with the extreme carboxy-terminal sequence, forms a central shelf-like structure. A C-terminal C2 domain mediates initial, Ca(2+)-dependent membrane binding. Most unexpectedly, however, electron microscopy reveals that the orientation of the perforin MACPF domain in the pore is inside-out relative to the subunit arrangement in CDCs. These data reveal remarkable flexibility in the mechanism of action of the conserved MACPF/CDC fold and provide new insights into how related immune defence molecules such as complement proteins assemble into pores.read more
Citations
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Journal ArticleDOI
Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores
Xing Liu,Zhibin Zhang,Zhibin Zhang,Jianbin Ruan,Jianbin Ruan,Youdong Pan,Venkat Giri Magupalli,Venkat Giri Magupalli,Hao Wu,Hao Wu,Judy Lieberman,Judy Lieberman +11 more
TL;DR: It is shown that GSDMD-NT oligomerizes in membranes to form pores that are visible by electron microscopy and kills cell-free bacteria in vitro and may have a direct bactericidal effect within the cytosol of host cells, but the importance of direct bacterial killing in controlling in vivo infection remains to be determined.
Journal ArticleDOI
Perforin and granzymes: function, dysfunction and human pathology
Ilia Voskoboinik,Ilia Voskoboinik,James C. Whisstock,James C. Whisstock,Joseph A. Trapani,Joseph A. Trapani +5 more
TL;DR: The current understanding of the structural, cellular and clinical aspects of perforin and granzyme biology is discussed, beginning to define and understand a range of human diseases that are associated with a failure to deliver active per forin to target cells.
Journal ArticleDOI
Pore-forming toxins: ancient, but never really out of fashion
TL;DR: The diverse pore architectures and membrane insertion mechanisms that have been revealed by structural studies of PFTs are discussed, and how these features contribute to binding specificity for different membrane targets are considered.
Journal ArticleDOI
Cryo-EM structure of the gasdermin A3 membrane pore
Jianbin Ruan,Jianbin Ruan,Shiyu Xia,Shiyu Xia,Xing Liu,Xing Liu,Judy Lieberman,Judy Lieberman,Hao Wu,Hao Wu +9 more
TL;DR: Cryo-electron microscopy structures of the 27-fold and 28-fold single-ring pores formed by the N-terminal fragment of mouse GSDMA3 (G SDMA3-NT) provide a basis that explains the activities of several mutant gasdermins, including defective mutants that are associated with cancer.
Journal ArticleDOI
Membrane Repair: Mechanisms and Pathophysiology
Sandra T. Cooper,Paul L. McNeil +1 more
TL;DR: Collective evidence reveals membrane repair employs primitive yet robust molecular machinery, such as vesicle fusion and contractile rings, processes evolutionarily honed for simplicity and success.
References
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Eric F. Pettersen,Thomas D. Goddard,Conrad C. Huang,Gregory S. Couch,Daniel M. Greenblatt,Elaine C. Meng,Thomas E. Ferrin +6 more
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