3D Crystallization and Structural Studies of Integral Membrane Proteins in Lipidic Cubic phases

Abstract Membrane protein research suffers from the drawback that detergents, which are commonly used to solubilize integral membrane proteins (IMPs), often lead to protein instability and reduced activity. Recently, lipid nanodiscs (NDs) and saposin-lipoprotein particles (Salipro) have emerged as alternative carrier systems that keep membrane proteins in a native-like lipidic solution environment and are suitable for biophysical and structural studies. Here, we systematically compare nanodiscs and Salipros with respect to long-term stability as well as activity and stability of the incorporated membrane protein using the ABC transporter MsbA as model system. Our results show that both systems are suitable for activity measurements as well as structural studies in solution. Based on our results we suggest screening of different lipids with respect to activity and stability of the incorporated IMP before performing structural studies.

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Overexpression of mammalian integral membrane proteins for structural studies

FEBS Letters ◽

10.1016/s0014-5793(01)02711-9 ◽

2001 ◽

Vol 504 (3) ◽

pp. 94-98 ◽

Cited By ~ 133

Author(s):

C.G. Tate

Keyword(s):

Membrane Proteins ◽

Integral Membrane Proteins ◽

Structural Studies

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A method for solution NMR structural studies of large integral membrane proteins: Reverse micelle encapsulation

Biochimica et Biophysica Acta (BBA) - Biomembranes ◽

10.1016/j.bbamem.2009.07.027 ◽

2010 ◽

Vol 1798 (2) ◽

pp. 150-160 ◽

Cited By ~ 23

Author(s):

Joseph M. Kielec ◽

Kathleen G. Valentine ◽

A. Joshua Wand

Keyword(s):

Membrane Proteins ◽

Reverse Micelle ◽

Integral Membrane Proteins ◽

Solution Nmr ◽

Structural Studies

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High-throughput analytical gel filtration screening of integral membrane proteins for structural studies

Biochimica et Biophysica Acta (BBA) - General Subjects ◽

10.1016/j.bbagen.2013.02.001 ◽

2013 ◽

Vol 1830 (6) ◽

pp. 3497-3508 ◽

Cited By ~ 22

Author(s):

Christian Löw ◽

Per Moberg ◽

Esben M. Quistgaard ◽

Marie Hedrén ◽

Fatma Guettou ◽

...

Keyword(s):

Membrane Proteins ◽

High Throughput ◽

Gel Filtration ◽

Integral Membrane Proteins ◽

Structural Studies

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Overexpression of integral membrane proteins for structural studies

Quarterly Reviews of Biophysics ◽

10.1017/s0033583500003504 ◽

1995 ◽

Vol 28 (3) ◽

pp. 315-422 ◽

Cited By ~ 286

Author(s):

R. Grisshammer ◽

C. G. Tateu

Keyword(s):

Membrane Proteins ◽

Light Harvesting ◽

Atomic Level ◽

Integral Membrane Proteins ◽

Biological Processes ◽

Structural Studies ◽

Light Harvesting Complex ◽

Prostaglandin H ◽

2D Crystals

Determination of the structure of integral membrane proteins is a challenging task that is essential to understand how fundamental biological processes (such as photosynthesis, respiration and solute translocation) function at the atomic level. Crystallisation of membrane proteins in 3D has led to the determination of four atomic resolution structures [photosynthetic reaction centres (Allenet al. 1987; Changet al. 1991; Deisenhofer & Michel, 1989; Ermleret al. 1994); porins (Cowanet al. 1992; Schirmeret al. 1995; Weisset al. 1991); prostaglandin H2synthase (Picotet al. 1994); light harvesting complex (McDermottet al. 1995)], and crystals of membrane proteins formed in the plane of the lipid bilayer (2D crystals) have produced two more structures [bacteriorhodopsin (Hendersonet al. 1990); light harvesting complex (Kühlbrandtet al. 1994)].

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