Purification of a membrane protein with conjugated engineered micelles

Guy Patchornik; Dganit Danino; Ellina Kesselman; Ellen Wachtel; Noga Friedman; Mordechai Sheves

doi:10.1021/bc400069w

Purification of a membrane protein with conjugated engineered micelles

Guy Patchornik^*, Dganit Danino, Ellina Kesselman, Ellen Wachtel, Noga Friedman, Mordechai Sheves

^*Corresponding author for this work

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

11 Scopus citations

Abstract

A novel method for purifying membrane proteins is presented. The approach makes use of engineered micelles composed of a nonionic detergent, β-octylglucoside, and a hydrophobic metal chelator, bathophenanthroline. Via the chelators, the micelles are specifically conjugated, i.e., tethered, in the presence of Fe²⁺ ions, thereby forming micellar aggregates which provide the environment for separation of lipid-soluble membrane proteins from water-soluble proteins. The micellar aggregates (here imaged by cryo-transmission electron microscopy) successfully purify the light driven proton pump, bacteriorhodopsin (bR), from E. coli lysate. Purification takes place within 15 min and can be performed both at room temperature and at 4 C. More than 94% of the water-soluble macromolecules in the lysate are excluded, with recovery yields of the membrane protein ranging between 74% and 85%. Since this approach does not require precipitants, high concentrations of detergent to induce micellar aggregates, high temperature, or changes in pH, it is suggested that it may be applied to the purification of a wide variety of membrane proteins.

Original language	English
Pages (from-to)	1270-1275
Number of pages	6
Journal	Bioconjugate Chemistry
Volume	24
Issue number	7
DOIs	https://doi.org/10.1021/bc400069w
State	Published - 17 Jul 2013
Externally published	Yes

Access to Document

10.1021/bc400069w

Cite this

@article{3e213417cdbd462896ff2c9911a4ae08,

title = "Purification of a membrane protein with conjugated engineered micelles",

abstract = "A novel method for purifying membrane proteins is presented. The approach makes use of engineered micelles composed of a nonionic detergent, β-octylglucoside, and a hydrophobic metal chelator, bathophenanthroline. Via the chelators, the micelles are specifically conjugated, i.e., tethered, in the presence of Fe2+ ions, thereby forming micellar aggregates which provide the environment for separation of lipid-soluble membrane proteins from water-soluble proteins. The micellar aggregates (here imaged by cryo-transmission electron microscopy) successfully purify the light driven proton pump, bacteriorhodopsin (bR), from E. coli lysate. Purification takes place within 15 min and can be performed both at room temperature and at 4 C. More than 94% of the water-soluble macromolecules in the lysate are excluded, with recovery yields of the membrane protein ranging between 74% and 85%. Since this approach does not require precipitants, high concentrations of detergent to induce micellar aggregates, high temperature, or changes in pH, it is suggested that it may be applied to the purification of a wide variety of membrane proteins.",

author = "Guy Patchornik and Dganit Danino and Ellina Kesselman and Ellen Wachtel and Noga Friedman and Mordechai Sheves",

year = "2013",

month = jul,

day = "17",

doi = "10.1021/bc400069w",

language = "英语",

volume = "24",

pages = "1270--1275",

journal = "Bioconjugate Chemistry",

issn = "1043-1802",

publisher = "American Chemical Society",

number = "7",

}

TY - JOUR

T1 - Purification of a membrane protein with conjugated engineered micelles

AU - Patchornik, Guy

AU - Danino, Dganit

AU - Kesselman, Ellina

AU - Wachtel, Ellen

AU - Friedman, Noga

AU - Sheves, Mordechai

PY - 2013/7/17

Y1 - 2013/7/17

N2 - A novel method for purifying membrane proteins is presented. The approach makes use of engineered micelles composed of a nonionic detergent, β-octylglucoside, and a hydrophobic metal chelator, bathophenanthroline. Via the chelators, the micelles are specifically conjugated, i.e., tethered, in the presence of Fe2+ ions, thereby forming micellar aggregates which provide the environment for separation of lipid-soluble membrane proteins from water-soluble proteins. The micellar aggregates (here imaged by cryo-transmission electron microscopy) successfully purify the light driven proton pump, bacteriorhodopsin (bR), from E. coli lysate. Purification takes place within 15 min and can be performed both at room temperature and at 4 C. More than 94% of the water-soluble macromolecules in the lysate are excluded, with recovery yields of the membrane protein ranging between 74% and 85%. Since this approach does not require precipitants, high concentrations of detergent to induce micellar aggregates, high temperature, or changes in pH, it is suggested that it may be applied to the purification of a wide variety of membrane proteins.

AB - A novel method for purifying membrane proteins is presented. The approach makes use of engineered micelles composed of a nonionic detergent, β-octylglucoside, and a hydrophobic metal chelator, bathophenanthroline. Via the chelators, the micelles are specifically conjugated, i.e., tethered, in the presence of Fe2+ ions, thereby forming micellar aggregates which provide the environment for separation of lipid-soluble membrane proteins from water-soluble proteins. The micellar aggregates (here imaged by cryo-transmission electron microscopy) successfully purify the light driven proton pump, bacteriorhodopsin (bR), from E. coli lysate. Purification takes place within 15 min and can be performed both at room temperature and at 4 C. More than 94% of the water-soluble macromolecules in the lysate are excluded, with recovery yields of the membrane protein ranging between 74% and 85%. Since this approach does not require precipitants, high concentrations of detergent to induce micellar aggregates, high temperature, or changes in pH, it is suggested that it may be applied to the purification of a wide variety of membrane proteins.

UR - http://www.scopus.com/inward/record.url?scp=84880394343&partnerID=8YFLogxK

U2 - 10.1021/bc400069w

DO - 10.1021/bc400069w

M3 - 文章

C2 - 23758098

AN - SCOPUS:84880394343

SN - 1043-1802

VL - 24

SP - 1270

EP - 1275

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 7

ER -

Purification of a membrane protein with conjugated engineered micelles

Abstract

Access to Document

Other files and links

Fingerprint

Cite this