Structure of a halophilic nucleoside diphosphate kinase from Halobacterium salinarum.
Nucleoside diphosphate kinase from the halophilic archaeon Halobacterium salinarum was crystallized in a free state and a substrate-bound form with CDP. The structures were solved to a resolution of 2.35 and 2.2A, respectively. Crystals with the apo-form were obtained with His6-tagged enzyme, whereas the untagged form was used for co-crystallization with the nucleotide. Crosslinking under different salt and pH conditions revealed a stronger oligomerization tendency for the tagged protein at low and high salt concentrations. The influence of the His6-tag on the halophilic nature of the enzyme is discussed on the basis of the observed structural properties.
Besir
Hüseyin
H
Max-Planck-Institut für Biochemie, Abteilung Membranbiochemie, Am Klopferspitz 18, 82152 Martinsried, Germany. hbesir@biochem.mpg.de
Zeth
Kornelius
K
Bracher
Andreas
A
Heider
Ursula
U
Ishibashi
Matsujiro
M
Tokunaga
Masao
M
Oesterhelt
Dieter
D
D003460:Crystallization; D018360:Crystallography, X-Ray; D003841:Deoxycytidine / Q000737:chemistry; D019281:Dimerization; D019617:Halobacterium salinarum / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry*; D009994:Osmolar Concentration; D011485:Protein Binding
D003841:Deoxycytidine; D009701:Nucleoside-Diphosphate Kinase