PRKRA Human

Protein Kinase IFN Double Stranded RNA Activator Human Recombinant
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Cat. No.
BT14443
Source
Escherichia Coli.
Synonyms
Interferon-inducible double-stranded RNA-dependent protein kinase activator A, Protein Kinase IFN Double Stranded RNA Activator, PKR-associated protein X, PKR-associating protein X, Protein activator of the interferon-induced protein kinase, Protein kinase, interferon-inducible double-stranded RNA-dependent activator, PRKRA, PACT, RAX, HSD-14, HSD14, Protein kinase, interferon-inducible double stranded RNA dependent activator, DYT16.
Appearance
Sterile Filtered colorless solution.
Purity
Greater than 80.0% as determined by SDS-PAGE.
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

Biological Function of PACT in Cellular Stress Response

PACT (Protein Activator of PKR) regulates stress-induced pathways via interaction with PKR (Protein Kinase R):

  1. PKR Activation: PACT binds to PKR, enabling its activation even in the absence of double-stranded RNA (dsRNA) .

  2. eIF2α Phosphorylation: Active PKR phosphorylates eIF2α, inhibiting translation elongation and triggering apoptosis under sustained stress .

  3. Interferon Signaling: PACT enhances interferon (IFN) production during viral infections, amplifying antiviral responses .

DYT-PRKRA Features:

  • Motor Symptoms: Generalized limb dystonia, laryngeal/neck involvement, parkinsonism .

  • Neurological Impact: Reduced Purkinje neuron dendritic arborization, striatal degeneration .

Luteolin as a Therapeutic Candidate

Luteolin reduces apoptosis in DYT-PRKRA cells by:

  • Modulating PACT-PKR Interaction: Inhibits constitutive PKR activation (5-fold higher in DYT-PRKRA vs. wild-type) .

  • Restoring Stress Response: Prevents excessive eIF2α phosphorylation and ER stress-induced cell death .

Murine Models of DYT-PRKRA

ModelKey FindingsSource
lear-5J MutationTruncated PACT → PKR inhibition → Cerebellar defects, dystonia
RAX KnockoutEmbryonic lethality, pituitary dysfunction

Interferon Dysregulation

DYT-PRKRA mutations may enhance IFN production, exacerbating neuroinflammation and post-febrile dystonia .

PRKRA Human in Experimental Research

PRKRA Human is utilized to study:

  1. Dystonia Mechanisms: In vitro models of PACT-PKR dysregulation in patient lymphoblasts .

  2. Apoptosis Pathways: Role of PKR-eIF2α axis in neurodegeneration .

  3. Drug Resistance: Overexpression linked to oxaliplatin resistance in mucinous ovarian cancer .

Product Specs

Introduction
Protein Kinase R (PKR) Activator, also known as PRKRA, is a protein involved in the cellular response to viral infections. Upon viral infection, PRKRA activates PKR even without the presence of double-stranded RNA (dsRNA), which is typically a hallmark of viral presence. This activation leads to the phosphorylation of eukaryotic initiation factor 2 alpha (eIF2α), ultimately inhibiting protein synthesis and triggering apoptosis, both of which are crucial for controlling viral spread. Mutations in the PRKRA gene have been linked to dystonia, a neurological disorder characterized by involuntary muscle contractions.
Description
This product consists of the human recombinant PRKRA protein, produced in E. coli. It is a single, non-glycosylated polypeptide chain composed of 336 amino acids (specifically, amino acids 1 through 313 of the native protein with an additional 23-amino acid Histidine tag at the N-terminus). The molecular weight of the recombinant protein is 36.8 kDa. The protein has undergone purification using proprietary chromatographic methods.
Physical Appearance
A clear and colorless solution that has been sterilized by filtration.
Formulation
The PRKRA protein is provided in a solution at a concentration of 0.25 mg/ml. The solution contains the following components: 20mM Tris-HCl buffer (pH 8.0), 0.2M NaCl, 50% glycerol, 2mM DTT, and 1mM EDTA.
Stability
For short-term storage (up to 4 weeks), the product can be stored at 4°C. For extended storage, it is recommended to freeze the product at -20°C. To ensure optimal stability during long-term storage, it is advisable to add a carrier protein such as albumin (HSA or BSA) to a final concentration of 0.1%. It is crucial to minimize repeated freeze-thaw cycles to maintain protein integrity.
Purity
The purity of the PRKRA protein is greater than 80%, as determined by SDS-PAGE analysis.
Synonyms
Interferon-inducible double-stranded RNA-dependent protein kinase activator A, Protein Kinase IFN Double Stranded RNA Activator, PKR-associated protein X, PKR-associating protein X, Protein activator of the interferon-induced protein kinase, Protein kinase, interferon-inducible double-stranded RNA-dependent activator, PRKRA, PACT, RAX, HSD-14, HSD14, Protein kinase, interferon-inducible double stranded RNA dependent activator, DYT16.
Source
Escherichia Coli.
Amino Acid Sequence
MGSSHHHHHH SSGLVPRGSH MGSMSQSRHR AEAPPLERED SGTFSLGKMI TAKPGKTPIQ VLHEYGMKTK NIPVYECERS DVQIHVPTFT FRVTVGDITC TGEGTSKKLA KHRAAEAAIN ILKANASICF AVPDPLMPDP SKQPKNQLNP IGSLQELAIH HGWRLPEYTL SQEGGPAHKR EYTTICRLES FMETGKGASK KQAKRNAAEK FLAKFSNISP ENHISLTNVV GHSLGCTWHS LRNSPGEKIN LLKRSLLSIP NTDYIQLLSE IAKEQGFNIT YLDIDELSAN GQYQCLAELS TSPITVCHGS GISCGNAQSD AAHNALQYLK IIAERK.

Q&A

Here’s a structured FAQ for researchers investigating PRKRA in human biology and disease, synthesized from peer-reviewed findings and genomic databases:

Advanced Research Questions

  • What experimental models resolve contradictions in PACT’s role in innate immunity?
    While PACT primarily activates PKR, some studies suggest it also regulates RIG-I and MDA-5 .

    • Resolution Strategy: Compare PKR-knockout vs. wild-type cells infected with VSV or SARS-CoV-2 to decouple PACT’s PKR-dependent and independent roles .

  • How does miR-122 modulate PRKRA expression, and what are the implications for viral replication?
    miR-122 represses PRKRA, enhancing HCV replication by reducing PKR-mediated antiviral signaling .

    • Experimental Design: Use luciferase reporters with PRKRA 3’UTR mutations to map miR-122 binding sites in hepatoma cells .

Technical Challenges & Solutions

  • What are the limitations of current PRKRA murine models for studying dystonia?
    Existing Prkra KO mice show neurodevelopmental deficits but lack robust dystonia phenotypes .

    • Innovative Approach: Generate conditional knock-in mice expressing human DYT16 mutations (e.g., P222L) in striatal neurons to mimic human pathology .

  • How can researchers differentiate PRKRA’s stress-specific roles (viral vs. ER stress)?

    • Multi-Omics Workflow:

      1. Transcriptomics (RNA-seq) of PRKRA-silenced cells under ER stress (thapsigargin) vs. viral mimic (poly(I:C)).

      2. Phosphoproteomics to quantify PKR and eIF2α activation states .

Data Interpretation Guidelines

  • Conflicting Apoptosis Results: Some studies report PACT as pro-apoptotic (via PKR), while others note anti-apoptotic roles in cancer. Context matters: cell type, stressor duration, and PACT expression levels (e.g., glioblastoma vs. neurons) .

  • Validation Tip: Cross-verify findings with PRKRA patient-derived iPSCs to ensure clinical relevance .

Product Science Overview

Introduction

Protein Kinase RNA-activated (PKR) is a crucial enzyme in the human body, encoded by the EIF2AK2 gene on chromosome 2 . It is a serine/threonine kinase that plays a significant role in the cellular response to viral infections and other stress signals. PKR is activated by double-stranded RNA (dsRNA), which is often produced during viral replication. This activation leads to the phosphorylation of the eukaryotic translation initiation factor 2 alpha (eIF2α), resulting in the inhibition of protein synthesis and the suppression of viral replication .

Mechanism of Action

PKR is primarily activated by binding to dsRNA, which induces its autophosphorylation and subsequent activation. Once activated, PKR phosphorylates eIF2α, leading to a reduction in protein synthesis. This mechanism is a part of the cell’s antiviral defense, as it helps to limit the production of viral proteins . Additionally, PKR can interact with other cellular proteins, such as PACT (Protein Activator of the Interferon-induced Protein Kinase), which further modulates its activity .

Role in Viral Infections

PKR plays a pivotal role in the cellular response to viral infections. For instance, during HIV-1 replication, PKR is transiently activated, leading to the phosphorylation of eIF2α and a decrease in viral protein synthesis . However, HIV-1 has evolved mechanisms to counteract this response. The virus can modulate the activity of PKR through various viral and cellular factors, including the interaction with PACT and ADAR1 (Adenosine Deaminase Acting on RNA), which can inhibit PKR activity and promote viral replication .

Therapeutic Potential

Given its crucial role in the antiviral response, PKR is a potential target for therapeutic interventions. Modulating PKR activity could enhance the body’s ability to fight viral infections. For example, enhancing PKR activation could improve the antiviral response, while inhibiting PKR could be beneficial in conditions where excessive PKR activity leads to detrimental effects, such as in certain inflammatory diseases .

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