Dataset Information

Photoinduced interactions of two dirhodium complexes with d(GTCGAC)2 probed by 2D NOESY.

ABSTRACT: The interactions between the 6-mer duplex oligonucleotide d(GTCGAC)2 and the photoactive dirhodium complexes cis-H,H-[Rh2(HNOCCH3)2(L)(CH3CN)4](2+), where L represents bpy (1, 2,2'-bipyridine) and dppz (2, dipyrido[3,2-a:2',3'-c]phenazine), were probed using 2D (1)H-(1)H NOESY NMR spectroscopy. Complex does not interact with the duplex in the dark, but binds covalently to the terminal guanine following irradiation with visible light. Similar behavior was observed for 2, but in addition to the photoinduced covalent DNA binding, the planar dppz ligand of the complex shields the terminal cytosine protons after irradiation. The results are consistent with photoinduced guanine coordination and end-capping of the duplex through ?-stacking interactions with the terminal GC base pair. These data show that in the presence of the 6-mer duplex oligonucleotide, 1 and 2 exhibit photoinduced covalent binding to DNA. In addition, the ?-stacking interactions of 2 with the duplex are enhanced upon irradiation.

SUBMITTER: Palmer AM

PROVIDER: S-EPMC4349562 | biostudies-other | 2015 Feb

REPOSITORIES: biostudies-other

ACCESS DATA

Publications

Photoinduced interactions of two dirhodium complexes with d(GTCGAC)2 probed by 2D NOESY.

Palmer Alycia M AM Knoll Jessica D JD Turro Claudia C

Dalton transactions (Cambridge, England : 2003) 20150201 8

The interactions between the 6-mer duplex oligonucleotide d(GTCGAC)2 and the photoactive dirhodium complexes cis-H,H-[Rh2(HNOCCH3)2(L)(CH3CN)4](2+), where L represents bpy (1, 2,2'-bipyridine) and dppz (2, dipyrido[3,2-a:2',3'-c]phenazine), were probed using 2D (1)H-(1)H NOESY NMR spectroscopy. Complex does not interact with the duplex in the dark, but binds covalently to the terminal guanine following irradiation with visible light. Similar behavior was observed for 2, but in addition to the ph ...[more]

PMID: 25557067

Similar Datasets

Project description:BackgroundHere we describe a novel approach used to identify the constituents of protein complexes with high fidelity, using the integrin-associated scaffolding protein PINCH as a test case. PINCH is comprised of five LIM domains, zinc-finger protein interaction modules. In Drosophila melanogaster, PINCH has two known high-affinity binding partners-Integrin-linked kinase (ILK) that binds to LIM1 and Ras Suppressor 1 (RSU1) that binds to LIM5-but has been postulated to bind additional proteins as well.ResultsTo purify PINCH complexes, in parallel we fused different affinity tags (Protein A and Flag) to different locations within the PINCH sequence (N- and C-terminus). We expressed these tagged versions of PINCH both in cell culture (overexpressed in Drosophila S2 cell culture in the presence of endogenous PINCH) and in vivo (at native levels in Drosophila lacking endogenous PINCH). After affinity purification, we analyzed PINCH complexes by a novel 2D-gel electrophoresis analysis, iGEO (interactions by 2D Gel Electrophoresis Overlap), with mass spectrometric identification of individual spots of interest. iGEO allowed the identification of protein partners that associate with PINCH under two independent purification strategies, providing confidence in the significance of the interaction. Proteins identified by iGEO were validated against a highly inclusive list of candidate PINCH interacting proteins identified in previous analyses by MuDPIT mass spectrometry.ConclusionsThe iGEO strategy confirmed a core complex comprised of PINCH, RSU1, ILK, and ILK binding partner Parvin. Our iGEO method also identified five novel protein partners that specifically interacted with PINCH in Drosophila S2 cell culture. Because of the improved reproducibility of 2D-GE methodology and the increasing affordability of the required labeling reagents, iGEO is a method that is accessible to most moderately well-equipped biological laboratories. The biochemical co-purifications inherent in iGEO allow for rapid and unambiguous identification of the constituents of protein complexes, without the need for extensive follow-up experiments.

Dataset Information

Photoinduced interactions of two dirhodium complexes with d(GTCGAC)2 probed by 2D NOESY.

Publications

Photoinduced interactions of two dirhodium complexes with d(GTCGAC)2 probed by 2D NOESY.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure