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Protein Expr Purif. 2004 Oct;37(2):479-85.

Ni-chelate-affinity purification and crystallization of the yeast mitochondrial F1-ATPase.

Author information

  • 1Department of Biochemistry and Molecular Biology, The Chicago Medical School, USA. David.Mueller@rosalindfranklin.edu

Abstract

The yeast mitochondrial ATPase has been genetically modified to include a His(6) Ni-affinity tag on the amino end of the mature beta-subunit. The modified beta-subunit is imported into the mitochondrion, properly processed to the mature form, and assembled into a mature and fully active ATP synthase. The F(1)-ATPase has been purified from submitochondrial particles after release from the membrane with chloroform, followed by Ni-chelate-affinity and gel filtration chromatography. The final enzyme is a homogeneous preparation with full activity and no apparent degradation products. This enzyme preparation has been used to obtain crystals that diffract to better than 2.8 A resolution.

PMID:
15358374
[PubMed - indexed for MEDLINE]
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