The structural basis for membrane binding and pore formation by lymphocyte perforin

Ruby H P Law, Natalya Lukoyanova, Ilia Voskoboinik, Tom T Caradoc-Davies, Katherine Baran, Michelle A Dunstone, Michael D'Angelo, Elena V Orlova, Fasseli J Coulibaly, Sandra Verschoor, Kylie A Browne, Annette Ciccone, Michael J Kuiper, Phillip I Bird, Joseph A Trapani, Helen R Saibil, James C Whisstock

Research output: Contribution to journalArticleResearchpeer-review

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 cell1a??6. 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)7. 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 a??key-shapeda?? molecule, comprising an amino-terminal membrane attack complex perforinlike (MACPF)/cholesterol dependent cytolysin (CDC) domain8,9 followed by an epidermal growth factor (EGF) domain that, together with the extreme carboxy-terminal sequence, forms a central shelf-like structure.AC-terminalC2domainmediates initial, Ca21-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 CDCs10,11. These data reveal remarkable flexibility in themechanism 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.
Original languageEnglish
Pages (from-to)447 - 451
Number of pages5
JournalNature
Volume468
Issue number7322
DOIs
Publication statusPublished - 2010

Cite this

Law, Ruby H P ; Lukoyanova, Natalya ; Voskoboinik, Ilia ; Caradoc-Davies, Tom T ; Baran, Katherine ; Dunstone, Michelle A ; D'Angelo, Michael ; Orlova, Elena V ; Coulibaly, Fasseli J ; Verschoor, Sandra ; Browne, Kylie A ; Ciccone, Annette ; Kuiper, Michael J ; Bird, Phillip I ; Trapani, Joseph A ; Saibil, Helen R ; Whisstock, James C. / The structural basis for membrane binding and pore formation by lymphocyte perforin. In: Nature. 2010 ; Vol. 468, No. 7322. pp. 447 - 451.
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title = "The structural basis for membrane binding and pore formation by lymphocyte perforin",
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 cell1a??6. 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)7. 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 a??key-shapeda?? molecule, comprising an amino-terminal membrane attack complex perforinlike (MACPF)/cholesterol dependent cytolysin (CDC) domain8,9 followed by an epidermal growth factor (EGF) domain that, together with the extreme carboxy-terminal sequence, forms a central shelf-like structure.AC-terminalC2domainmediates initial, Ca21-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 CDCs10,11. These data reveal remarkable flexibility in themechanism 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.",
author = "Law, {Ruby H P} and Natalya Lukoyanova and Ilia Voskoboinik and Caradoc-Davies, {Tom T} and Katherine Baran and Dunstone, {Michelle A} and Michael D'Angelo and Orlova, {Elena V} and Coulibaly, {Fasseli J} and Sandra Verschoor and Browne, {Kylie A} and Annette Ciccone and Kuiper, {Michael J} and Bird, {Phillip I} and Trapani, {Joseph A} and Saibil, {Helen R} and Whisstock, {James C}",
year = "2010",
doi = "10.1038/nature09518",
language = "English",
volume = "468",
pages = "447 -- 451",
journal = "Nature",
issn = "0028-0836",
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Law, RHP, Lukoyanova, N, Voskoboinik, I, Caradoc-Davies, TT, Baran, K, Dunstone, MA, D'Angelo, M, Orlova, EV, Coulibaly, FJ, Verschoor, S, Browne, KA, Ciccone, A, Kuiper, MJ, Bird, PI, Trapani, JA, Saibil, HR & Whisstock, JC 2010, 'The structural basis for membrane binding and pore formation by lymphocyte perforin' Nature, vol. 468, no. 7322, pp. 447 - 451. https://doi.org/10.1038/nature09518

The structural basis for membrane binding and pore formation by lymphocyte perforin. / Law, Ruby H P; Lukoyanova, Natalya; Voskoboinik, Ilia; Caradoc-Davies, Tom T; Baran, Katherine; Dunstone, Michelle A; D'Angelo, Michael; Orlova, Elena V; Coulibaly, Fasseli J; Verschoor, Sandra; Browne, Kylie A; Ciccone, Annette; Kuiper, Michael J; Bird, Phillip I; Trapani, Joseph A; Saibil, Helen R; Whisstock, James C.

In: Nature, Vol. 468, No. 7322, 2010, p. 447 - 451.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - The structural basis for membrane binding and pore formation by lymphocyte perforin

AU - Law, Ruby H P

AU - Lukoyanova, Natalya

AU - Voskoboinik, Ilia

AU - Caradoc-Davies, Tom T

AU - Baran, Katherine

AU - Dunstone, Michelle A

AU - D'Angelo, Michael

AU - Orlova, Elena V

AU - Coulibaly, Fasseli J

AU - Verschoor, Sandra

AU - Browne, Kylie A

AU - Ciccone, Annette

AU - Kuiper, Michael J

AU - Bird, Phillip I

AU - Trapani, Joseph A

AU - Saibil, Helen R

AU - Whisstock, James C

PY - 2010

Y1 - 2010

N2 - 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 cell1a??6. 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)7. 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 a??key-shapeda?? molecule, comprising an amino-terminal membrane attack complex perforinlike (MACPF)/cholesterol dependent cytolysin (CDC) domain8,9 followed by an epidermal growth factor (EGF) domain that, together with the extreme carboxy-terminal sequence, forms a central shelf-like structure.AC-terminalC2domainmediates initial, Ca21-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 CDCs10,11. These data reveal remarkable flexibility in themechanism 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.

AB - 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 cell1a??6. 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)7. 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 a??key-shapeda?? molecule, comprising an amino-terminal membrane attack complex perforinlike (MACPF)/cholesterol dependent cytolysin (CDC) domain8,9 followed by an epidermal growth factor (EGF) domain that, together with the extreme carboxy-terminal sequence, forms a central shelf-like structure.AC-terminalC2domainmediates initial, Ca21-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 CDCs10,11. These data reveal remarkable flexibility in themechanism 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.

UR - http://www.nature.com/nature/journal/v468/n7322/pdf/nature09518.pdf

U2 - 10.1038/nature09518

DO - 10.1038/nature09518

M3 - Article

VL - 468

SP - 447

EP - 451

JO - Nature

JF - Nature

SN - 0028-0836

IS - 7322

ER -