1 research outputs found
Interaction of the HIV NCp7 Protein with Platinum(II) and Gold(III) Complexes Containing Tridentate Ligands
The human immunodeficiency
virus (HIV) nucleocapsid protein (NCp7) plays significant roles in
the virus life cycle and has been targeted by compounds that could
lead to its denaturation or block its interaction with viral RNA.
Herein, we describe the interactions of platinumÂ(II) and goldÂ(III)
complexes with NCp7 and how the reactivity/affinity of potential inhibitors
can be modulated by judicious choice of ligands. The interactions
of [MClÂ(N<sub>3</sub>)]<sup><i>n</i>+</sup> (M = Pt<sup>2+</sup> (<i>n</i> = 1) and Au<sup>3+</sup> (<i>n</i> = 2); N<sub>3</sub> = tridentate chelate ligands: bisÂ(2-pyridylmethyl)Âmethylamine
(Mebpma, <b>L</b><sup><b>1</b></sup>) and bisÂ(2-pyridylmethyl)Âamine
(bpma, <b>L</b><sup><b>2</b></sup>) with the C-terminal
zinc finger of NCp7 (ZF2) were investigated by electrospray ionization-mass
spectroscopy (ESI-MS). Mass spectra from the incubation of [MClÂ(Mebpma)]<sup><i>n</i>+</sup> complexes (<b>PtL</b><sup><b>1</b></sup> and <b>AuL</b><sup><b>1</b></sup>) with ZF2 indicated
that they were more reactive than the previously studied diethylenetriamine-containing
analogues [MClÂ(dien)]<sup><i>n</i>+</sup>. The initial product
of reaction of <b>PtL</b><sup><b>1</b></sup> with ZF2
results in loss of all ligands and release of zinc to give the platinated
apopeptide {PtF} (F = apopeptide). This is in contrast to the incubation
with [PtClÂ(dien)]<sup>+</sup>, in which {PtÂ(dien)}–peptide
adducts are observed. Incubation of the Au<sup>3+</sup> complex <b>AuL</b><sup><b>1</b></sup> with ZF2 gave Au<sub><i>x</i></sub>F<sup><i>n</i>+</sup> species (<i>x</i> = 1, 2, 4, F = apopeptide) again with loss of all ligands.
Furthermore, the formally substitution-inert analogues [PtÂ(N<sub>3</sub>)ÂL]<sup>2+</sup> (L = 4-methylpyridine (4-pic), 4-dimethylaminopyridine
(dmap), and 9-ethylguanine (9-EtGua)) were prepared to examine stacking
interactions with <i>N</i>-acetyltryptophan (N-AcTrp), the
Trp-containing ZF2, and the “full” two-finger NCp7 itself
using fluorescence quenching titration. Use of bpma and Mebpma gave
slightly higher affinity than analogous [PtÂ(dien)ÂL)]<sup>2+</sup> complexes.
The dmap-containing complexes (<b>PtL</b><sup><b>1</b></sup><b>a</b> and <b>PtL</b><sup><b>2</b></sup><b>a</b>) had the greatest association constants (<i>K</i><sub>a</sub>) for N-AcTrp and ZF2 peptide. The complex <b>PtL</b><sup><b>1</b></sup><b>a</b> had the highest <i>K</i><sub>a</sub> when compared with other known Pt<sup>2+</sup> analogues:
[PtÂ(dien)Â(9-EtGua)]<sup>2+</sup> < [PtÂ(bpma)Â(9-EtGua)]<sup>2+</sup> < [PtÂ(dien)Â(dmap)]<sup>2+</sup>< <b>PtL</b><sup><b>2</b></sup><b>a</b> < <b>PtL</b><sup><b>1</b></sup><b>a</b>. A <i>K</i><sub>a</sub> value
of ca. 40.6 ± 1.0 × 10<sup>3</sup> M<sup>–1</sup> was obtained for the full NCp7 peptide with <b>PtL</b><sup><b>1</b></sup><b>a</b>. In addition, the mass spectrum
of the interaction between ZF2 and <b>PtL</b><sup><b>1</b></sup><b>a</b> confirms formation of a 1:1 <b>PtL</b><sup><b>1</b></sup><b>a</b>/ZF2 adduct. The reactivity
of selected complexes with sulfur-containing amino acid <i>N</i>-acetylcysteine (N-AcCys) was also investigated by <sup>195</sup>Pt and <sup>1</sup>H NMR spectroscopy and ESI-MS. The precursor compounds
[PtClÂ(N<sub>3</sub>)]<sup>+</sup> <b>PtL</b><sup><b>1</b></sup> and <b>PtL</b><sup><b>2</b></sup> reacted readily,
whereas their [PtÂ(N<sub>3</sub>)ÂL]<sup>2+</sup> analogues <b>PtL</b><sup><b>1</b></sup><b>a</b> and <b>PtL</b><sup><b>2</b></sup><b>a</b> were inert to substitution