High-Resolution Solution NMR Structure of the Minimal Active Domain of the Human Immunodeficiency Virus Type-2 Nucleocapsid Protein
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- 作者：Yoshio Kodera ; Kazuki Sato ; Tomonori Tsukahara ; Hiroyoshi Komatsu ; Tadakazu Maeda ; and Toshiyuki Kohno
- 刊名：Biochemistry
- 出版年：1998
- 出版时间：December 22, 1998
- 年：1998
- 卷：37
- 期：51
- 页码：17704 - 17713
- 全文大小：222K
- 年卷期：v.37,no.51(December 22, 1998)
- ISSN：1520-4995
文摘
The retroviral nucleocapsid (NC) protein is a multifunctional protein essential for RNA genomepackaging and viral infectivity. The NC protein, NCp8, of the human immunodeficiency virus type-II(HIV-2) is a 49 amino acid peptide containing two zinc fingers, of the type C-X2-C-X4-H-X4-C, connectedby seven amino acid residues, called the "basic amino acid cluster." It has been shown that the N-terminalzinc finger flanked by the basic amino acid cluster is the minimal active domain for the specific bindingto viral RNA and other functions. However, the structure-activity relationships of NCp8 have not beeninvestigated in detail. In the present study, the three-dimensional structure of a 29 amino acid peptide,including the minimal active domain (NCp8-f1), was determined by two-dimensional 1H NMR spectroscopywith simulated annealing calculations. A total of 15 converged structures of NCp8-f1 were obtained onthe basis of 355 experimental constraints, including 343 distance constraints obtained from nuclearOverhauser effect connectivities, 12 torsion angle (
, 
1) constraints, and four constraints for zinc binding.The root-mean-square deviation of the 15 converged structures was 0.29 ± 0.04 Å for the backboneatoms (N, C
, C) and 1.27 ± 0.13 Å for all heavy atoms. Interestingly, the basic amino acid cluster itselfwas defined well, with a loop-like conformation in which three arginine residues in the cluster and onearginine residue in the zinc finger are located approximately in the same plane of the molecule and areexposed to the solvent. The structure-activity relationships are discussed on the basis of the comparisonof this well-defined structure with those of other NC proteins.
, 1) constraints, and four constraints for zinc binding.The root-mean-square deviation of the 15 converged structures was 0.29 ± 0.04 Å for the backboneatoms (N, C, C) and 1.27 ± 0.13 Å for all heavy atoms. Interestingly, the basic amino acid cluster itselfwas defined well, with a loop-like conformation in which three arginine residues in the cluster and onearginine residue in the zinc finger are located approximately in the same plane of the molecule and areexposed to the solvent. The structure-activity relationships are discussed on the basis of the comparisonof this well-defined structure with those of other NC proteins.