Biology Reference
In-Depth Information
CHAPTER
13
Mem brane Reconstitu tion
OUTLINE
A. Detergents
266
Vesicles by Detergent Dilution
289
Membrane Detergents
269
LUVs: Dilution Methods
290
Concentration Methods
291
B. Membrane Protein Isolation
271
E. Membrane Reconstitution
292
C. Membrane Lipid Isolation
275
Ca 2รพ ATPase
293
TLC
276
Bacterial Proline Transporter
294
HPLC
278
Fatty Acid Analysis
279
F. Lipid Rafts
295
Experimental Support of Rafts
297
D. Model Lipid Bilayer Membranes
e
Questions Concerning Rafts
298
Liposomes
282
One Last Word on Rafts
299
Model Membranes
282
Lipid Raft Characteristics
299
Liposomes
284
Unilamellar Vesicles
287
Summary
300
SUV by Sonication
288
References
300
Vesicles by Extrusion
288
Membranes are unimaginably complex. They consist of hundreds of different proteins
floating in a bewildering and crowded sea of a thousand or more different lipids and this
mixture is in constant flux. A membrane is not homogeneous but exists in fleeting patches
that exhibit both lateral and trans-membrane asymmetry. Also, countless reactions occur
simultaneously in and around the membrane. Can the biological membrane problem be
simplified? In this chapter we will examine how one membrane protein can be isolated
away from the others and reconstituted into a model lipid bilayer membrane in order to
determine how the protein functions. Finally we will investigate the isolation and properties
of an important membrane domain involved in cell signaling
the lipid raft.
e
 
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