Two-step purification of outer membrane proteins

K Beis; C Whitfield; IR Booth; James Henderson Naismith

doi:10.1016/j.ijbiomac.2005.12.008

Two-step purification of outer membrane proteins

K Beis, C Whitfield, IR Booth, James Henderson Naismith

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

18 Citations (Scopus)

Abstract

Here, we describe a simple and efficient method for the purification of Escherichia coli outer membrane proteins. We have tested this protocol for the purification of Wza and Osmoporin C (OmpC) proteins. Both proteins were purified to homogeneity, in two steps, by anion exchange and size exclusion chromatography with a final yield of 92.5 mg for the Wza protein and 291.5 mg for the OmpC protein. The purity of the samples was judged by electrophoretic analysis, mass spectrometry, single particle analysis, three-dimensional (3D) crystallisation and X-ray diffraction. (c) 2005 Elsevier B.V. All rights reserved.

Original language	English
Pages (from-to)	10-14
Number of pages	5
Journal	International Journal of Biological Macromolecules
Volume	39
Issue number	1-3
DOIs	https://doi.org/10.1016/j.ijbiomac.2005.12.008
Publication status	Published - 15 Aug 2006

Keywords

membrane protein purification
Wza
OmpC
OmpA
MALDI-TOF
electron microscopy
3D crystals
X-ray diffraction
GROUP-1 CAPSULAR POLYSACCHARIDE
ESCHERICHIA-COLI
TRANSLOCATION
WZA
CRYSTALLIZATION
LIPOPROTEIN
EXPRESSION
CRYSTALS
SURFACE
PORIN

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10.1016/j.ijbiomac.2005.12.008

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title = "Two-step purification of outer membrane proteins",

abstract = "Here, we describe a simple and efficient method for the purification of Escherichia coli outer membrane proteins. We have tested this protocol for the purification of Wza and Osmoporin C (OmpC) proteins. Both proteins were purified to homogeneity, in two steps, by anion exchange and size exclusion chromatography with a final yield of 92.5 mg for the Wza protein and 291.5 mg for the OmpC protein. The purity of the samples was judged by electrophoretic analysis, mass spectrometry, single particle analysis, three-dimensional (3D) crystallisation and X-ray diffraction. (c) 2005 Elsevier B.V. All rights reserved.",

keywords = "membrane protein purification, Wza, OmpC, OmpA, MALDI-TOF, electron microscopy, 3D crystals, X-ray diffraction, GROUP-1 CAPSULAR POLYSACCHARIDE, ESCHERICHIA-COLI, TRANSLOCATION, WZA, CRYSTALLIZATION, LIPOPROTEIN, EXPRESSION, CRYSTALS, SURFACE, PORIN",

author = "K Beis and C Whitfield and IR Booth and Naismith, {James Henderson}",

year = "2006",

month = aug,

day = "15",

doi = "10.1016/j.ijbiomac.2005.12.008",

language = "English",

volume = "39",

pages = "10--14",

journal = "International Journal of Biological Macromolecules",

issn = "0141-8130",

publisher = "Elsevier",

number = "1-3",

}

TY - JOUR

T1 - Two-step purification of outer membrane proteins

AU - Beis, K

AU - Whitfield, C

AU - Booth, IR

AU - Naismith, James Henderson

PY - 2006/8/15

Y1 - 2006/8/15

N2 - Here, we describe a simple and efficient method for the purification of Escherichia coli outer membrane proteins. We have tested this protocol for the purification of Wza and Osmoporin C (OmpC) proteins. Both proteins were purified to homogeneity, in two steps, by anion exchange and size exclusion chromatography with a final yield of 92.5 mg for the Wza protein and 291.5 mg for the OmpC protein. The purity of the samples was judged by electrophoretic analysis, mass spectrometry, single particle analysis, three-dimensional (3D) crystallisation and X-ray diffraction. (c) 2005 Elsevier B.V. All rights reserved.

AB - Here, we describe a simple and efficient method for the purification of Escherichia coli outer membrane proteins. We have tested this protocol for the purification of Wza and Osmoporin C (OmpC) proteins. Both proteins were purified to homogeneity, in two steps, by anion exchange and size exclusion chromatography with a final yield of 92.5 mg for the Wza protein and 291.5 mg for the OmpC protein. The purity of the samples was judged by electrophoretic analysis, mass spectrometry, single particle analysis, three-dimensional (3D) crystallisation and X-ray diffraction. (c) 2005 Elsevier B.V. All rights reserved.

KW - membrane protein purification

KW - Wza

KW - OmpC

KW - OmpA

KW - MALDI-TOF

KW - electron microscopy

KW - 3D crystals

KW - X-ray diffraction

KW - GROUP-1 CAPSULAR POLYSACCHARIDE

KW - ESCHERICHIA-COLI

KW - TRANSLOCATION

KW - WZA

KW - CRYSTALLIZATION

KW - LIPOPROTEIN

KW - EXPRESSION

KW - CRYSTALS

KW - SURFACE

KW - PORIN

UR - http://www.scopus.com/inward/record.url?scp=33745991802&partnerID=8YFLogxK

U2 - 10.1016/j.ijbiomac.2005.12.008

DO - 10.1016/j.ijbiomac.2005.12.008

M3 - Article

SN - 0141-8130

VL - 39

SP - 10

EP - 14

JO - International Journal of Biological Macromolecules

JF - International Journal of Biological Macromolecules

IS - 1-3

ER -

Two-step purification of outer membrane proteins

Abstract

Keywords

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