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Isolation, crystallization and crystal structure determination of bovine kidney Na + ,K + ‐ATPase
Author(s) -
Gregersen Jonas Lindholt,
Mattle Daniel,
Fedosova Natalya U.,
Nissen Poul,
Reinhard Linda
Publication year - 2016
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x1600279x
Subject(s) - crystallization , crystallography , orthorhombic crystal system , crystal structure , chemistry , atpase , molecular replacement , ouabain , sodium , enzyme , biochemistry , organic chemistry
Na + ,K + ‐ATPase is responsible for the transport of Na + and K + across the plasma membrane in animal cells, thereby sustaining vital electrochemical gradients that energize channels and secondary transporters. The crystal structure of Na + ,K + ‐ATPase has previously been elucidated using the enzyme from native sources such as porcine kidney and shark rectal gland. Here, the isolation, crystallization and first structure determination of bovine kidney Na + ,K + ‐ATPase in a high‐affinity E2–BeF 3 − –ouabain complex with bound magnesium are described. Crystals belonging to the orthorhombic space group C 222 1 with one molecule in the asymmetric unit exhibited anisotropic diffraction to a resolution of 3.7 Å with full completeness to a resolution of 4.2 Å. The structure was determined by molecular replacement, revealing unbiased electron‐density features for bound BeF 3 − , ouabain and Mg 2+ ions.