PKACa2- RIa2
Inactive Protein Kinase A holoenzyme type I alpha Recombinant
Cat. No.
BT8009
Source
Escherichia Coli.
Synonyms
Protein Kinase A holoenzyme type I alpha, PKACa2- RIa2.
Appearance
Sterile Filtered clear solution.
Purity
Greater than 95.0% as determined by SDS-PAGE.
Usage
Prospec'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
Inactive holoenzyme consisting of one dimeric regulatory subunit type I alpha and two monomeric catalytic subunits (cAMP-free).
Protein Kinase A Recombinant is purified by proprietary chromatographic techniques.
Protein Kinase A Recombinant is purified by proprietary chromatographic techniques.
Product Specs
Introduction
Protein kinase A (PKA) is a crucial enzyme that plays a central role in various cellular processes. Its holoenzyme structure consists of two catalytic subunits responsible for enzymatic activity and two regulatory subunits that modulate this activity. The catalytic subunits contain domains for ATP binding, regulatory subunit interaction, and the active site. Conversely, the regulatory subunits, existing as dimers, possess cyclic AMP (cAMP) binding domains, catalytic subunit interaction domains, and an auto-inhibitory domain. These regulatory subunits exhibit diversity, with type I and type II, each further classified into alpha and beta subtypes, encoded by distinct genes and exhibiting differential cellular and tissue distribution. Type I enzymes are predominantly cytoplasmic, while type II enzymes tend to associate with cellular membranes. This intricate interplay between catalytic and regulatory subunits underscores the complexity of PKA regulation and its significance in cellular signaling.
Description
This product consists of the inactive PKA holoenzyme, specifically the type I alpha isoform. It comprises a dimeric regulatory subunit (RIα) bound to two monomeric catalytic subunits. This holoenzyme is in its cAMP-free state, indicating inactivity. The recombinant protein has been purified using proprietary chromatographic techniques to ensure high purity.
Physical Appearance
The product is a clear solution that has undergone sterile filtration.
Formulation
The PKA holoenzyme type-I alpha is supplied in a solution containing 50% glycerol to maintain its stability.
Stability
To ensure product stability, store PKA at 4°C if you plan to use the entire vial within 2-4 weeks. For long-term storage, storing the product at -20°C is recommended. Repeated freezing and thawing cycles should be avoided to prevent protein degradation.
Purity
Analysis by SDS-PAGE reveals that the purity of this product is greater than 95%.
Biological Activity
The inactive PKA holoenzyme can be activated by introducing the second messenger cAMP, which binds to the regulatory subunits. The activation constant for this process is approximately 100nM. Upon activation, the two monomeric catalytic subunits are released, enabling the enzyme to phosphorylate its substrates.
Synonyms
Protein Kinase A holoenzyme type I alpha, PKACa2- RIa2.
Source
Escherichia Coli.
Product Science Overview
Structure and Composition
The inactive PKA holoenzyme type I alpha consists of:
- One dimeric regulatory subunit type I alpha: This subunit is responsible for binding cyclic adenosine monophosphate (cAMP) and regulating the activity of the catalytic subunits.
- Two monomeric catalytic subunits: These subunits are responsible for the enzyme’s catalytic activity but remain inactive when bound to the regulatory subunits in the absence of cAMP .
Mechanism of Activation
Purification and Stability
The recombinant form of the inactive PKA holoenzyme type I alpha is typically expressed in Escherichia coli (E. coli) and purified using proprietary chromatographic techniques. The protein is supplied in a sterile filtered clear solution, often formulated with 50% glycerol to enhance stability .
For storage, it is recommended to keep the protein at -20°C for long-term use, while short-term storage at 4°C is suitable if the entire vial will be used within 2-4 weeks. It is essential to avoid multiple freeze-thaw cycles to maintain the protein’s integrity .
Applications and Significance
The inactive PKA holoenzyme type I alpha recombinant is valuable in various research applications, including:
- Studying cAMP signaling pathways: Understanding the role of PKA in cellular processes and how it is regulated by cAMP.
- Drug discovery and development: Screening for compounds that can modulate PKA activity, which may have therapeutic potential for diseases related to dysregulated PKA signaling.
- Biotechnological applications: Utilizing the recombinant protein in assays and experiments to explore its properties and interactions with other molecules.
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