Histone Acetyltransferase/Deacetylase Activity Assay Service

Q: Q: What types of acetylation events can your assays detect?

A: Our assays cover mono-acetylation on lysine residues (e.g., H3K9ac, H3K14ac) and can be adapted for multi-acetylation events. We tailor detection methods based on the specific modification and research needs.

Q: Q: Do you offer services for drug discovery applications?

A: Absolutely. Our services include inhibitor screening, lead optimization, and mechanistic studies tailored to drug discovery pipelines, with support for ADME-Tox integration.

Q: Q: Can your assays distinguish between different HDAC classes?

A: Yes, we employ class-specific inhibitors, substrate profiling, and kinetic analyses to differentiate between HDAC classes (e.g., Class I vs. III sirtuins).

Inquiry

CD BioSciences offers specialized histone acetyltransferase (HAT) and deacetylase (HDAC) activity assay services to enable accurate profiling of enzymatic functions in epigenetic regulation. Our platform integrates advanced methodologies for quantifying acetylation dynamics, supporting research in gene expression, disease mechanisms, and therapeutic development. We provide end-to-end solutions tailored to diverse needs, from basic kinetic studies to high-throughput inhibitor screening.

Introduction to Histone Acetyltransferase and Deacetylase

Histone acetylation and deacetylation are pivotal epigenetic modifications that regulate chromatin structure and gene transcription. HATs catalyze the addition of acetyl groups to lysine residues on histone tails, promoting an open chromatin state and facilitating transcriptional activation. Conversely, HDACs remove these acetyl groups, leading to chromatin condensation and transcriptional repression.

Feature	Histone Acetyltransferases (HATs)	Histone Deacetylases (HDACs)
Enzyme Classification	Type A: Nuclear, acetylates chromatin-bound histones (e.g., p300/CBP, GNAT, MYST families). Type B: Cytoplasmic, acetylates newly synthesized free histones.	Class I (HDAC1, 2, 3, 8) Class II (IIa: HDAC4, 5, 7, 9; IIb: HDAC6, 10) Class III (Sirtuins 1-7) Class IV (HDAC11)
Effect on Gene Expression	Activates transcription. The open chromatin allows transcription factors and RNA polymerase to bind more easily, promoting gene expression.	Represses transcription. The condensed chromatin hinders the binding of the transcription machinery, leading to gene silencing.
Key Regulators & Inhibitors	Activity is influenced by the availability of its substrate, acetyl-CoA, which links it directly to cellular metabolic status.	HDAC Inhibitors (HDACis) are a class of epigenetic drugs. Examples include Vorinostat and Romidepsin, which are approved for treating certain lymphomas.
Biological Significance	Acts as a transcriptional co-activator. Dysregulation is associated with developmental disorders and cancers.	Often overexpressed in cancers, leading to silencing of tumor suppressor genes. HDAC inhibitors can reactivate latent viruses by disrupting viral gene silencing.

Significance of Histone Acetyltransferase/Deacetylase Activity Assay

Assaying HAT and HDAC activity are essential for elucidating epigenetic mechanisms, screening for potential therapeutic agents (such as HAT agonists or HDAC inhibitors), and identifying disease biomarkers. In drug discovery, high-throughput screening platforms using these assays are instrumental in developing novel anticancer drugs like HDAC inhibitors (e.g., Vorinostat). Furthermore, in basic research, activity assays help decipher the "histone code" by quantifying modifications at specific sites (e.g., H3K9ac, H3K14ac) and understanding their roles in processes like stem cell differentiation and responses to environmental stimuli.

The mechanism of methylation and demethylation on the R group nitrogen of the histone lysine tail.

Fig.1 Schematic representation of histone acetylation and deacetylation process. (Miziak P, et al., 2024)

Our Services

CD BioSciences delivers customized assay solutions for characterizing HAT/HDAC activities, leveraging robust protocols and state-of-the-art detection technologies. Our services are designed to provide high sensitivity, specificity, and reproducibility.

Core Detection Methodologies

Fluorescence-Based Assays

Utilize fluorogenic substrates (e.g., acetylated peptides with fluorescent tags) for high-throughput applications with minimal background.

Luminescence Detection

Leverage luciferase-reporting systems for enhanced sensitivity in low-abundance enzyme assays.

Radioisotopic Methods

Employ labeled substrates (e.g., ³H-acetyl-CoA) for direct, quantitative detection of acetylation changes.

Mass Spectrometry

Provide precise quantification of acetylation states with site-specific resolution, ideal for complex samples.

Samples Requirements

To ensure optimal assay performance, we provide detailed guidelines for sample preparation and submission.

Enzyme Samples: Purified HATs/HDACs with >90% purity, specified concentration, storage buffer, and known activity references.
Substrate Preparations: Histone proteins, peptides, or nucleosomes with defined modifications (e.g., acetylation states), purity >95%, and concentration details.
Biological Samples: Cell lysates or tissue extracts with information on preparation method, protein concentration, and storage conditions.
Compound Libraries: For screening, provide compound structures, concentrations, and solvent compatibility data.

Deliverables

Upon project completion, you will receive:

A detailed report summarizing experimental procedures, raw data (e.g., kinetic curves, dose-response plots), and analytical results.
Validation data confirming assay specificity, including control experiments and reproducibility metrics.
Expert interpretation highlighting key findings, such as inhibitor potency, substrate preferences, or mechanistic insights.
Recommendations for follow-up studies, such as cellular validation or structural analyses.

Frequently Asked Questions (FAQs)

Q: What types of acetylation events can your assays detect?

Q: Do you offer services for drug discovery applications?

Q: Can your assays distinguish between different HDAC classes?

Reference

1. Miziak P, Baran M, Borkiewicz L, et al. Acetylation of histone H3 in cancer progression and prognosis[J]. International Journal of Molecular Sciences, 2024, 25(20): 10982.