Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2021946 | Protein Expression and Purification | 2006 | 7 Pages |
Abstract
The cytoplasmic, N-terminal domain (Nt) of the electrogenic sodium/bicarbonate cotransporter-NBCe1-over-expresses in Escherichia coli and yields a large amount of soluble protein. A novel purification strategy, which involves a streptomycin precipitation, overcomes obstacles of instability and copurifying proteins, and leads to the first seen Nt-NBCe1 crystals. The purification procedure generally lends itself to the purification of Nts from other classes of the SLC4 family. Size-exclusion chromatography suggests that the Nt of NBCe1 as well as the Nt of other SLC4 members form dimers. A comparison of Nt-NBCe1 to SLC4 member Nt-AE1, based on purification properties and predicted secondary-structure sequence alignments, suggests a similar mechanism for dimer stabilization.
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Authors
Harindarpal S. Gill, Walter F. Boron,