Expression, refolding and crystallization of the OpcA invasin from Neisseria meningitidis

S. M. Prince; C. Feron; D. Janssens; Y. Lobet; M. Achtman; B. Kusecek; P. A. Bullough; J. P. Derrick

doi:10.1107/S0907444901009416

Expression, refolding and crystallization of the OpcA invasin from Neisseria meningitidis

S. M. Prince, C. Feron, D. Janssens, Y. Lobet, M. Achtman, B. Kusecek, P. A. Bullough, J. P. Derrick

Research output: Contribution to journal › Article › peer-review

Abstract

OpcA is an integral outer membrane from the Gram-negative pathogen Neisseria meningitidis that plays a role in adhesion of meningococci to host cells. The protein was overexpressed in Escherichia coli in an insoluble form and a procedure developed for refolding by rapid dilution from denaturant into detergent solution. The refolded material was identical to native OpcA isolated from meningococci, as judged by overall molecular weight, migration on SDS-PAGE and reaction against monoclonal antibodies. Both native and recombinant OpcA crystallized under similar conditions to give an orthorhombic crystal form (P21212), with unit-cell parameters a = 96.9, b = 46.3, c = 74.0 Å. Complete data sets of reflections were collected from native and refolded OpcA to 2.0 Å resolution.

Original language	English
Pages (from-to)	1164-1166
Number of pages	2
Journal	Acta Crystallographica Section D: Biological Crystallography
Volume	57
Issue number	8
DOIs	https://doi.org/10.1107/S0907444901009416
Publication status	Published - 2001

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title = "Expression, refolding and crystallization of the OpcA invasin from Neisseria meningitidis",

abstract = "OpcA is an integral outer membrane from the Gram-negative pathogen Neisseria meningitidis that plays a role in adhesion of meningococci to host cells. The protein was overexpressed in Escherichia coli in an insoluble form and a procedure developed for refolding by rapid dilution from denaturant into detergent solution. The refolded material was identical to native OpcA isolated from meningococci, as judged by overall molecular weight, migration on SDS-PAGE and reaction against monoclonal antibodies. Both native and recombinant OpcA crystallized under similar conditions to give an orthorhombic crystal form (P21212), with unit-cell parameters a = 96.9, b = 46.3, c = 74.0 {\AA}. Complete data sets of reflections were collected from native and refolded OpcA to 2.0 {\AA} resolution.",

author = "Prince, {S. M.} and C. Feron and D. Janssens and Y. Lobet and M. Achtman and B. Kusecek and Bullough, {P. A.} and Derrick, {J. P.}",

year = "2001",

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language = "English",

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publisher = "International Union of Crystallography",

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TY - JOUR

T1 - Expression, refolding and crystallization of the OpcA invasin from Neisseria meningitidis

AU - Prince, S. M.

AU - Feron, C.

AU - Janssens, D.

AU - Lobet, Y.

AU - Achtman, M.

AU - Kusecek, B.

AU - Bullough, P. A.

AU - Derrick, J. P.

PY - 2001

Y1 - 2001

N2 - OpcA is an integral outer membrane from the Gram-negative pathogen Neisseria meningitidis that plays a role in adhesion of meningococci to host cells. The protein was overexpressed in Escherichia coli in an insoluble form and a procedure developed for refolding by rapid dilution from denaturant into detergent solution. The refolded material was identical to native OpcA isolated from meningococci, as judged by overall molecular weight, migration on SDS-PAGE and reaction against monoclonal antibodies. Both native and recombinant OpcA crystallized under similar conditions to give an orthorhombic crystal form (P21212), with unit-cell parameters a = 96.9, b = 46.3, c = 74.0 Å. Complete data sets of reflections were collected from native and refolded OpcA to 2.0 Å resolution.

AB - OpcA is an integral outer membrane from the Gram-negative pathogen Neisseria meningitidis that plays a role in adhesion of meningococci to host cells. The protein was overexpressed in Escherichia coli in an insoluble form and a procedure developed for refolding by rapid dilution from denaturant into detergent solution. The refolded material was identical to native OpcA isolated from meningococci, as judged by overall molecular weight, migration on SDS-PAGE and reaction against monoclonal antibodies. Both native and recombinant OpcA crystallized under similar conditions to give an orthorhombic crystal form (P21212), with unit-cell parameters a = 96.9, b = 46.3, c = 74.0 Å. Complete data sets of reflections were collected from native and refolded OpcA to 2.0 Å resolution.

U2 - 10.1107/S0907444901009416

DO - 10.1107/S0907444901009416

M3 - Article

C2 - 11468407

SN - 0907-4449

VL - 57

SP - 1164

EP - 1166

JO - Acta Crystallographica Section D: Biological Crystallography

JF - Acta Crystallographica Section D: Biological Crystallography

IS - 8

ER -

Expression, refolding and crystallization of the OpcA invasin from Neisseria meningitidis

Abstract

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