Recombinant Human GKAP1 Protein: Properties & Applications

Recombinant Human GKAP1 Protein is a cGMP-dependent protein kinase anchoring protein produced recombinantly for use in biochemical assays, protein interaction studies, and investigation of cGMP-mediated signaling pathways.

Protein Name

G Kinase Anchoring Protein 1 (GKAP1)

Also Known As

cGMP-dependent protein kinase anchoring protein, GKAP

Species Origin

Human (Homo sapiens)

Protein Function

Anchors cGMP-dependent protein kinase (PKG) to specific subcellular compartments, facilitating spatially regulated cGMP signaling

Expression System

Recombinant expression system (specific host system; see product datasheet)

Primary Research Applications

Protein-protein interaction assays, cGMP signaling studies, kinase anchoring studies, Western blot, ELISA

SKU

rp375060

What Is GKAP1 and Why Does It Matter?

GKAP1, or G Kinase Anchoring Protein 1, is a member of the A-kinase anchoring protein (AKAP) superfamily that specifically interacts with cGMP-dependent protein kinase (PKG, also termed protein kinase G). By tethering PKG to defined subcellular locations, GKAP1 ensures that cGMP signals are transduced with high spatial and temporal precision. This subcellular targeting is essential for the regulation of diverse physiological processes, including smooth muscle relaxation, platelet aggregation, cardiac function, and neuronal signaling.

Unlike many AKAPs that anchor cAMP-dependent protein kinase (PKA), GKAP1 shows selectivity for the cGMP/PKG axis, making it a uniquely valuable target for researchers dissecting the molecular mechanisms of nitric oxide (NO) and natriuretic peptide signaling cascades. Dysregulation of cGMP signaling has been implicated in cardiovascular disease, pulmonary hypertension, and neurological disorders, highlighting the pathophysiological relevance of studying GKAP1 interactions.

Biochemical Properties of Recombinant Human GKAP1 Protein

The Recombinant Human GKAP1 Protein (SKU: rp375060) is produced using a recombinant expression platform to ensure consistent batch-to-batch quality and biological activity. Researchers should consult the product datasheet for confirmed details on molecular weight, purity grade, formulation buffer, storage conditions, and recommended reconstitution procedures, as these parameters are critical for experimental reproducibility.

Key quality attributes typically characterized for recombinant proteins of this class include:

• Purity: Assessed by SDS-PAGE and/or HPLC to confirm the absence of significant contaminants.
• Biological Activity: Validated through binding assays confirming interaction with PKG or relevant binding partners.
• Formulation: Lyophilized or liquid formulation in a stabilizing buffer; specific details are product-lot dependent.
• Storage: Typically stored at -20°C or -80°C; repeated freeze-thaw cycles should be avoided to maintain protein integrity.

Always refer to the certificate of analysis (CoA) supplied with each lot for lot-specific quality data.

Research Applications of GKAP1 Protein

Recombinant Human GKAP1 Protein is well-suited for a range of in vitro research applications focused on cGMP signaling and kinase anchoring biology:

• Protein–Protein Interaction Studies: GKAP1 can be used as bait or prey in pull-down assays, co-immunoprecipitation (co-IP), or surface plasmon resonance (SPR) experiments to map its interactome, especially its interaction with PKG isoforms.
• Binding and Competition Assays: Biochemical assays using recombinant GKAP1 can quantify binding affinities and identify small molecules or peptides that disrupt the GKAP1–PKG interaction, supporting drug discovery efforts.
• Western Blot and ELISA Standards: The recombinant protein serves as a positive control or standard curve antigen in immunoassays detecting endogenous GKAP1.
• Structural and Biophysical Studies: Recombinant GKAP1 can support crystallography, NMR, or cryo-EM studies aimed at elucidating the structural basis of kinase anchoring.
• Cell-Free Reconstitution: Used to reconstitute cGMP signaling complexes in vitro for mechanistic studies outside the complexity of intact cells.

These applications make Recombinant Human GKAP1 Protein a versatile reagent for cardiovascular, neurological, and cell-signaling research programs.

Handling, Storage, and Quality Considerations

To ensure optimal performance of Recombinant Human GKAP1 Protein in downstream applications, researchers should adhere to the following best practices:

• Reconstitution: If supplied in lyophilized form, reconstitute with sterile, ultrapure water or the buffer recommended in the datasheet. Allow the vial to equilibrate to room temperature before opening.
• Aliquoting: Prepare single-use aliquots immediately after reconstitution to minimize protein degradation from repeated freeze-thaw cycles.
• Storage Temperature: Store at -80°C for long-term stability. Short-term storage at -20°C is acceptable for periods specified on the datasheet.
• Avoid Detergents and Extreme pH: Non-ionic detergents and pH extremes can denature the protein and compromise activity; follow buffer guidelines provided in the CoA.
• Carrier Protein: For very low concentration working solutions, consider adding a carrier protein (e.g., BSA) as specified in the application protocol to prevent adsorptive losses.

Lot-specific purity, activity, and endotoxin data are provided in the CoA. Contact Aladdin Scientific technical support for application-specific guidance or custom formulation requests.

Frequently asked questions

What is the biological function of GKAP1?

GKAP1 (G Kinase Anchoring Protein 1) functions as a scaffold protein that anchors cGMP-dependent protein kinase (PKG) to specific subcellular compartments. This spatial localization enables precise, localized transduction of cGMP signals generated downstream of nitric oxide and natriuretic peptide receptor activation.

Which expression system is used to produce Recombinant Human GKAP1 Protein (rp375060)?

The specific recombinant expression host system for rp375060 is detailed in the product datasheet. Please refer to the datasheet or contact Aladdin Scientific technical support for confirmed expression system information, as this can affect protein glycosylation status and activity in some assay formats.

How should I store Recombinant Human GKAP1 Protein to maintain its activity?

For long-term storage, keep the protein at -80°C. Avoid repeated freeze-thaw cycles by preparing single-use aliquots upon reconstitution. Refer to the lot-specific CoA for exact storage buffer composition, recommended working concentration, and shelf-life information.

Can Recombinant Human GKAP1 Protein be used as a positive control in Western blot experiments?

Yes, Recombinant Human GKAP1 Protein is suitable for use as a positive control antigen in Western blot and ELISA experiments. It provides a well-defined band at the expected molecular weight and can be used to validate antibody specificity and assay sensitivity when detecting endogenous GKAP1 in cell or tissue lysates.

Is Recombinant Human GKAP1 Protein suitable for drug discovery screening assays?

Recombinant Human GKAP1 Protein is well-suited for in vitro drug discovery applications, particularly for identifying small molecules or peptides that disrupt the GKAP1–PKG protein–protein interaction. It can be incorporated into fluorescence polarization, SPR, or AlphaScreen-based binding assays. Confirm assay compatibility with the product datasheet specifications before designing your screening campaign.

References

1. Welch EJ, Jones BW, Scott JD. Networking with AKAPs: context-dependent regulation of anchored enzymes. Mol Interv. 2010;10(2):86-97.
2. Francis SH, Busch JL, Corbin JD, Sibley D. cGMP-dependent protein kinases and cGMP phosphodiesterases in nitric oxide and cGMP action. Pharmacol Rev. 2010;62(3):525-563.
3. Aladdin Scientific. Recombinant Human GKAP1 Protein, SKU rp375060. Product Datasheet. Aladdin Scientific; [current version].

Products are supplied for research and development use only. Not for use in humans, animals, diagnosis, or therapy.