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Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G

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Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G. / Atkins, Angela; Wyborn, Neil R.; Wallace, Alistair J.; Stillman, Timothy J.; Black, Lance K.; Fielding, Andrew B.; Hisakado, Masataka; Artymiuk, Peter J.; Green, Jeffrey.

In: Journal of Biological Chemistry, Vol. 275, No. 52, 29.12.2000, p. 41150-41155.

Research output: Contribution to Journal/MagazineJournal articlepeer-review

Harvard

Atkins, A, Wyborn, NR, Wallace, AJ, Stillman, TJ, Black, LK, Fielding, AB, Hisakado, M, Artymiuk, PJ & Green, J 2000, 'Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G', Journal of Biological Chemistry, vol. 275, no. 52, pp. 41150-41155. https://doi.org/10.1074/jbc.M005420200

APA

Atkins, A., Wyborn, N. R., Wallace, A. J., Stillman, T. J., Black, L. K., Fielding, A. B., Hisakado, M., Artymiuk, P. J., & Green, J. (2000). Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G. Journal of Biological Chemistry, 275(52), 41150-41155. https://doi.org/10.1074/jbc.M005420200

Vancouver

Atkins A, Wyborn NR, Wallace AJ, Stillman TJ, Black LK, Fielding AB et al. Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G. Journal of Biological Chemistry. 2000 Dec 29;275(52):41150-41155. https://doi.org/10.1074/jbc.M005420200

Author

Atkins, Angela ; Wyborn, Neil R. ; Wallace, Alistair J. ; Stillman, Timothy J. ; Black, Lance K. ; Fielding, Andrew B. ; Hisakado, Masataka ; Artymiuk, Peter J. ; Green, Jeffrey. / Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 52. pp. 41150-41155.

Bibtex

@article{f83312b7359a4b48a6807dd165a13be9,
title = "Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G",
abstract = "The novel pore-forming toxin hemolysin E (HlyE, ClyA, or SheA) consists of a long four-helix bundle with a subdomain (beta tongue) that interacts with target membranes at one pole and an additional helix (alpha(G)) that, with the four long helices, forms a five-helix bundle (tail domain) at the other pole. Random amino acid substitutions that impair hemolytic activity were clustered mostly, but not exclusively, within the tail domain, specifically amino acids within, adjacent to, or interacting with alpha(G). Deletion of amino acids downstream of alpha(G) did not affect activity, but deletions encompassing alpha(G) yielded insoluble and inactive proteins. In the periplasm Cys-285 (alpha(G)) is linked to Cys-87 (alpha(B)) of the four-helix bundle via an intramolecular disulfide. Oxidized HlyE did not form spontaneously in vitro but could be generated by addition of Cu(II) or mimicked by treatment with Hg(II) salts to yield inactive proteins. Such treatments did not affect binding to target membranes nor assembly into non-covalently linked octameric complexes once associated with a membrane. However, gel filtration analyses suggested that immobilizing alpha(G) inhibits oligomerization in solution. Thus once associated with a membrane, immobilizing alpha(G) inhibits HlyE activity at a late stage of pore formation, whereas in solution it prevents aggregation and consequent inactivation.",
keywords = "Amino Acid Sequence, Hemolysin Proteins, Molecular Sequence Data, Molecular Weight, Protein Structure, Secondary, Structure-Activity Relationship, Journal Article, Research Support, Non-U.S. Gov't",
author = "Angela Atkins and Wyborn, {Neil R.} and Wallace, {Alistair J.} and Stillman, {Timothy J.} and Black, {Lance K.} and Fielding, {Andrew B.} and Masataka Hisakado and Artymiuk, {Peter J.} and Jeffrey Green",
year = "2000",
month = dec,
day = "29",
doi = "10.1074/jbc.M005420200",
language = "English",
volume = "275",
pages = "41150--41155",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "52",

}

RIS

TY - JOUR

T1 - Structure-function relationships of a novel bacterial toxin, hemolysin E. The role of alpha G

AU - Atkins, Angela

AU - Wyborn, Neil R.

AU - Wallace, Alistair J.

AU - Stillman, Timothy J.

AU - Black, Lance K.

AU - Fielding, Andrew B.

AU - Hisakado, Masataka

AU - Artymiuk, Peter J.

AU - Green, Jeffrey

PY - 2000/12/29

Y1 - 2000/12/29

N2 - The novel pore-forming toxin hemolysin E (HlyE, ClyA, or SheA) consists of a long four-helix bundle with a subdomain (beta tongue) that interacts with target membranes at one pole and an additional helix (alpha(G)) that, with the four long helices, forms a five-helix bundle (tail domain) at the other pole. Random amino acid substitutions that impair hemolytic activity were clustered mostly, but not exclusively, within the tail domain, specifically amino acids within, adjacent to, or interacting with alpha(G). Deletion of amino acids downstream of alpha(G) did not affect activity, but deletions encompassing alpha(G) yielded insoluble and inactive proteins. In the periplasm Cys-285 (alpha(G)) is linked to Cys-87 (alpha(B)) of the four-helix bundle via an intramolecular disulfide. Oxidized HlyE did not form spontaneously in vitro but could be generated by addition of Cu(II) or mimicked by treatment with Hg(II) salts to yield inactive proteins. Such treatments did not affect binding to target membranes nor assembly into non-covalently linked octameric complexes once associated with a membrane. However, gel filtration analyses suggested that immobilizing alpha(G) inhibits oligomerization in solution. Thus once associated with a membrane, immobilizing alpha(G) inhibits HlyE activity at a late stage of pore formation, whereas in solution it prevents aggregation and consequent inactivation.

AB - The novel pore-forming toxin hemolysin E (HlyE, ClyA, or SheA) consists of a long four-helix bundle with a subdomain (beta tongue) that interacts with target membranes at one pole and an additional helix (alpha(G)) that, with the four long helices, forms a five-helix bundle (tail domain) at the other pole. Random amino acid substitutions that impair hemolytic activity were clustered mostly, but not exclusively, within the tail domain, specifically amino acids within, adjacent to, or interacting with alpha(G). Deletion of amino acids downstream of alpha(G) did not affect activity, but deletions encompassing alpha(G) yielded insoluble and inactive proteins. In the periplasm Cys-285 (alpha(G)) is linked to Cys-87 (alpha(B)) of the four-helix bundle via an intramolecular disulfide. Oxidized HlyE did not form spontaneously in vitro but could be generated by addition of Cu(II) or mimicked by treatment with Hg(II) salts to yield inactive proteins. Such treatments did not affect binding to target membranes nor assembly into non-covalently linked octameric complexes once associated with a membrane. However, gel filtration analyses suggested that immobilizing alpha(G) inhibits oligomerization in solution. Thus once associated with a membrane, immobilizing alpha(G) inhibits HlyE activity at a late stage of pore formation, whereas in solution it prevents aggregation and consequent inactivation.

KW - Amino Acid Sequence

KW - Hemolysin Proteins

KW - Molecular Sequence Data

KW - Molecular Weight

KW - Protein Structure, Secondary

KW - Structure-Activity Relationship

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1074/jbc.M005420200

DO - 10.1074/jbc.M005420200

M3 - Journal article

C2 - 11006277

VL - 275

SP - 41150

EP - 41155

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 52

ER -