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Nat Protoc. 2009;4(5):619-37. doi: 10.1038/nprot.2009.27.

A general protocol for the crystallization of membrane proteins for X-ray structural investigation.

Author information

1
Department of Biochemistry and Biophysics, University of California in San Francisco, 600 16th Street, San Francisco, California 94158-2517, USA.

Abstract

Protein crystallography is used to generate atomic resolution structures of protein molecules. These structures provide information about biological function, mechanism and interaction of a protein with substrates or effectors including DNA, RNA, cofactors or other small molecules, ions and other proteins. This technique can be applied to membrane proteins resident in the membranes of cells. To accomplish this, membrane proteins first need to be either heterologously expressed or purified from a native source. The protein has to be extracted from the lipid membrane with a mild detergent and purified to a stable, homogeneous population that may then be crystallized. Protein crystals are then used for X-ray diffraction to yield atomic resolution structures of the desired membrane protein target. Below, we present a general protocol for the growth of diffraction quality membrane protein crystals. The process of protein crystallization is highly variable, and obtaining diffraction quality crystals can require weeks to months or even years in some cases.

PMID:
19360018
PMCID:
PMC4075773
DOI:
10.1038/nprot.2009.27
[Indexed for MEDLINE]
Free PMC Article

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