Déméné H, Jullian N, Morellet N, de Rocquigny H, Cornille F, Maigret B, Roques B P
Départemente de Pharmacochimie Moléculaire et Structurale, U266 INSERM-URA D 1500 CNRS, Faculté de Pharmacie, Paris, France.
J Biomol NMR. 1994 Mar;4(2):153-70. doi: 10.1007/BF00175244.
The nucleocapsid protein of Moloney murine leukemia virus (NCp10) is a 56-amino acid protein which contains one zinc finger of the CysX2CysX4HisX4Cys form, a highly conserved motif present in most retroviruses and retroelements. At pH > or = 5, NCp10 binds one zinc atom and the complexation induces a folding of the CysX2CysX4HEsX4Cys box, similar to that observed for the zinc-binding domains of HIV-1 NC protein. The three-dimensional structure of NCp10 has been determined in aqueous solution by 600 MHz 1H NMR spectroscopy. The proton resonances could be almost completely assigned by means of phase-sensitive double-quantum-filtered COSY, TOCSY and NOESY techniques. NOESY spectra yielded 597 relevant structural constraints, which were used as input for distance geometry calculations with DIANA. Further refinement was performed by minimization with the program AMBER, which was modified by introducing a zinc force field. The solution structure is characterized by a well-defined central zinc finger (rmsd of 0.747 +/- 0.209 A for backbone atoms and 1.709 +/- 0.187 A when all atoms are considered), surrounded by flexible N- and C-terminal domains. The Tyr28, Trp35, Lys37, Lys41 and Lys42 residues, which are essential for activity, lie on the same face of the zinc finger, forming a bulge structure probably involved in viral RNA binding. The significance of these structural characteristics for the various biological functions of the protein is discussed, taking into account the results obtained with various mutants.
莫洛尼鼠白血病病毒的核衣壳蛋白（NCp10）是一种由56个氨基酸组成的蛋白质，它含有一个CysX2CysX4HisX4Cys形式的锌指结构，这是大多数逆转录病毒和逆转录元件中存在的高度保守基序。在pH≥5时，NCp10结合一个锌原子，这种络合诱导CysX2CysX4HEsX4Cys盒发生折叠，类似于在HIV-1 NC蛋白的锌结合结构域中观察到的情况。通过600 MHz 1H NMR光谱法在水溶液中确定了NCp10的三维结构。质子共振几乎可以通过相敏双量子滤波COSY、TOCSY和NOESY技术完全归属。NOESY光谱产生了597个相关的结构约束，这些约束被用作使用DIANA进行距离几何计算的输入。通过使用引入锌力场进行修改的AMBER程序进行最小化进一步优化。溶液结构的特征是有一个明确的中央锌指（主链原子的均方根偏差为0.747±0.209 Å，考虑所有原子时为1.709±0.187 Å），周围是灵活的N端和C端结构域。对活性至关重要的Tyr28、Trp35、Lys37、Lys41和Lys42残基位于锌指的同一面上，形成一个可能参与病毒RNA结合的凸起结构。考虑到各种突变体获得的结果，讨论了这些结构特征对该蛋白质各种生物学功能的意义。
