1EHW
HUMAN NUCLEOSIDE DIPHOSPHATE KINASE 4
1EHW の概要
エントリーDOI | 10.2210/pdb1ehw/pdb |
分子名称 | NUCLEOSIDE DIPHOSPHATE KINASE, SULFATE ION (3 entities in total) |
機能のキーワード | nucleoside diphosphate kinase, nm23, mitochondrial, killer-of-prune, transferase |
由来する生物種 | Homo sapiens (human) |
細胞内の位置 | Mitochondrion intermembrane space ; Peripheral membrane protein: O00746 |
タンパク質・核酸の鎖数 | 2 |
化学式量合計 | 36707.62 |
構造登録者 | Milon, L.,Meyer, P.,Chiadmi, M.,Munier, A.,Johansson, M.,Karlsson, A.,Lascu, I.,Capeau, J.,Janin, J.,Lacombe, M.-L. (登録日: 2000-02-23, 公開日: 2000-05-17, 最終更新日: 2024-02-07) |
主引用文献 | Milon, L.,Meyer, P.,Chiadmi, M.,Munier, A.,Johansson, M.,Karlsson, A.,Lascu, I.,Capeau, J.,Janin, J.,Lacombe, M.L. The human nm23-H4 gene product is a mitochondrial nucleoside diphosphate kinase. J.Biol.Chem., 275:14264-14272, 2000 Cited by PubMed Abstract: We demonstrate here the catalytic activity and subcellular localization of the Nm23-H4 protein, product of nm23-H4, a new member of the human nm23/nucleoside diphosphate (NDP) kinase gene family (Milon, L., Rousseau-Merck, M., Munier, A., Erent, M., Lascu, I., Capeau, J., and Lacombe, M. L. (1997) Hum. Genet. 99, 550-557). Nm3-H4 was synthesized in escherichia coli as the full-length protein and as a truncated form missing the N-terminal extension characteristic of mitochondrial targeting. The truncated form possesses NDP kinase activity, whereas the full-length protein is inactive, suggesting that the extension prevents enzyme folding and/or activity. X-ray crystallographic analysis was performed on active truncated Nm23-H4. Like other eukaryotic NDP kinases, it is a hexamer. Nm23-H4 naturally possesses a serine residue at position 129, equivalent to the K-pn mutation of the Drosophila NDP kinase. The x-ray structure shows that the presence of Ser(129) has local structural effects that weaken subunit interactions. Site-directed mutagenesis shows that the serine is responsible for the lability of Nm23-H4 to heat and urea treatment, because the S129P mutant is greatly stabilized. Examination of human embryonic kidney 293 cells transfected with green fluorescent protein fusions by confocal microscopy shows a specific mitochondrial localization of Nm23-H4 that was also demonstrated by Western blot analysis of subcellular fractions of these cells. Import into mitochondria is accompanied by cleavage of the N-terminal extension that results in NDP kinase activity. Submitochondrial fractionation indicates that Nm23-H4 is associated with mitochondrial membranes, possibly to the contact sites between the outer and inner membranes. PubMed: 10799505DOI: 10.1074/jbc.275.19.14264 主引用文献が同じPDBエントリー |
実験手法 | X-RAY DIFFRACTION (2.4 Å) |
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