Identifying protein interactions with metal-modified DNA using microarray technology

Abstract

Protein microarrays have been used extensively to identify protein-protein interactions; however, this technology has not been widely applied to protein-DNA interactions. In particular, this work demonstrates the utility of this technique for rapidly identifying interactions of proteins with metal-modified DNA. Protein macroarray experiments were carried out with high mobility group protein 1 (HMG-1) and cisplatin- and chromium-modified 50-mer oligonucleotides to demonstrate "proof of principle." Commercially available protein microarrays containing many different classes of human proteins were then employed to search for additional interactions with cisplatin-modified DNA. The results of the microarray experiments confirmed some known interactions and, more importantly, identified many novel protein interactions, demonstrating the utility of this method as a rapid, high-throughput technique to discover proteins that interact with metal-modified DNA.

Chromium interactions in plants: current status and future strategies

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Protein microarrays: high-throughput tools for proteomics

“…This report also identified a novel DNA-binding protein with a metabolic enzyme activity directly regulating eukaryotic gene expression [72]. Recently, commercial protein microarrays have been used for the large-scale search of protein-DNA interactions [73]. A microarray composed of 282 known and predicted yeast transcription factors detected numerous pairings of transcription factors and their binding DNA sequences.…”

Protein microarray: A key approach of proteomics

“…Stansfield et al employed commercially available protein microarrays to screen additional interactions of cisplatinmodified DNA, and identified many novel protein-DNA interactions (Stansfield et al, 2009). A protein microarray containing 4256 proteins encoded from Escherichia coli K12 strain was used to develop assays for identifying proteins involved in the recognition of potential base damage in DNA .…”