Histones, the highly conserved proteins responsible for packaging DNA into chromatin, are subject to an array of chemical modifications. These post-translational modifications (PTMs) occur predominantly on their unstructured N-terminal "tail" regions and include methylation, acetylation, phosphorylation, and more. Each PTM can serve as a regulatory signal, influencing chromatin structure and gene expression. For instance, acetylation often correlates with gene activation, while methylation can have activating or repressive effects depending on the context and specific histone residue targeted. Next to DNA methylation, histone methylation and acetylation are the most well-characterized epigenetic marks. To investigate these modifications, researchers employ a variety of methods, one of which is histone peptide arrays.
What are Histone Peptide Arrays?
Histone peptide arrays are analytical tools that facilitate the systematic investigation of histone PTMs and their interactions with various biomolecules. They involve the immobilization of synthetic histone peptides, each carrying a specific PTM, onto a solid surface like a glass slide or a nitrocellulose membrane. In general, these peptides consist of a short amino acid sequence (typically comprising 10 to 20 amino acids) derived from the histone tail region, containing the PTM of interest. By arranging these peptides in an array format, researchers can simultaneously study the binding affinity and specificity of various proteins, antibodies, or enzymes for specific PTMs.
Applications
The scientific applications of histone peptide arrays in epigenetic research are wide-ranging. These applications not only advance our fundamental understanding of epigenetic regulation, but also hold great promise for drug discovery, biomarker identification, and therapeutic development in the context of various diseases:
Epigenetic Profiling: Histone peptide arrays are instrumental in profiling the binding preferences of epigenetic reader proteins, such as bromodomains, chromodomains, and tudor domains. These readers play pivotal roles in interpreting the histone code and recruiting various effector proteins involved in transcriptional regulation, DNA repair, and chromatin remodeling. This allows researchers to gain insights into the epigenetic code and how it regulates gene expression.
Drug Discovery: They play a crucial role in drug discovery and development, helping identify small molecules or compounds that can modulate specific PTMs, potentially leading to novel epigenetic therapies.
Biomarker Discovery: By studying the interactions between histone PTMs and reader proteins, researchers can identify potential biomarkers for various diseases, including cancer and neurological disorders.
Limitations and Advancements
While conventional histone peptide array methods offer valuable insights into epigenetic research, they are not without limitations: (a) time-consuming; (b) require highly specialized equipment; and, in particular, (c) have an insufficient variety of modified histones in the array. To address these shortcomings and advance the field of histone peptide arrays, EpigenTek has developed the Pre-Sure™ Histone H3 Peptide Array ELISA Kit. With this kit, selectivity and specificity of antibodies can be screened against the most comprehensive variety of histone H3 modifications (46 different PTMs), including the most important methylation (36), acetylation (8) and phosphorylation (2) modifications. Alternatively, the kit can be used for identifying histone modifying enzyme substrates or for analyzing specificity of histone binding proteins, providing flexibility according to your specific research needs. The kit also offers the following additional advantages:
Quick and efficient procedure that can be completed in under 2 hours.
High quality peptides (>95% pure) that are HPLC-purified and mass spec-verified.
Peptides are coated in 2 concentrations, which allows for the screening of both strong and weak reactive antibodies, enzymes, and histone binding proteins.
ELISA-based detection system, making the assay easy and convenient.
Simple, reliable, and consistent assay conditions.
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