3ZTP

Orthorhombic crystal form P21212 of the Aquifex aeolicus nucleoside diphosphate kinase

3ZTP の概要

• エントリーDOI: 10.2210/pdb3ztp/pdb
• 関連するPDBエントリー: 3ZTO 3ZTQ 3ZTR 3ZTS
• 分子名称: NUCLEOSIDE DIPHOSPHATE KINASE, GLYCEROL, SULFATE ION, ... (4 entities in total)
• 機能のキーワード: transferase
• 由来する生物種: AQUIFEX AEOLICUS
• 細胞内の位置: Cytoplasm (By similarity): O67528
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 32210.87

構造登録者

Boissier, F.,Georgescauld, F.,Moynie, L.,Dupuy, J.-W.,Sarger, C.,Podar, M.,Lascu, I.,Giraud, M.-F.,Dautant, A. (登録日: 2011-07-12, 公開日: 2012-03-14, 最終更新日: 2024-10-16)

主引用文献

Boissier, F.,Georgescauld, F.,Moynie, L.,Dupuy, J.W.,Sarger, C.,Podar, M.,Lascu, I.,Giraud, M.F.,Dautant, A.
An Intersubunit Disulfide Bridge Stabilizes the Tetrameric Nucleoside Diphosphate Kinase of Aquifex Aeolicus.
Proteins, 80:1658-, 2012

PubMed Abstract: The nucleoside diphosphate kinase (Ndk) catalyzes the reversible transfer of the γ-phosphate from nucleoside triphosphate to nucleoside diphosphate. Ndks form hexamers or two types of tetramers made of the same building block, namely, the common dimer. The secondary interfaces of the Type I tetramer found in Myxococcus xanthus Ndk and of the Type II found in Escherichia coli Ndk involve the opposite sides of subunits. Up to now, the few available structures of Ndk from thermophiles were hexameric. Here, we determined the X-ray structures of four crystal forms of the Ndk from the hyperthermophilic bacterium Aquifex aeolicus (Aa-Ndk). Aa-Ndk displays numerous features of thermostable proteins and is made of the common dimer but it is a tetramer of Type I. Indeed, the insertion of three residues in a surface-exposed spiral loop, named the Kpn-loop, leads to the formation of a two-turn α-helix that prevents both hexamer and Type II tetramer assembly. Moreover, the side chain of the cysteine at position 133, which is not present in other Ndk sequences, adopts two alternate conformations. Through the secondary interface, each one forms a disulfide bridge with the equivalent Cys133 from the neighboring subunit. This disulfide bridge was progressively broken during X-ray data collection by radiation damage. Such crosslinks counterbalance the weakness of the common-dimer interface. A 40% decrease of the kinase activity at 60°C after reduction and alkylation of the protein corroborates the structural relevance of the disulfide bridge on the tetramer assembly and enzymatic function.

PubMed: 22467275
DOI: 10.1002/PROT.24062

実験手法

X-RAY DIFFRACTION (1.37 Å)