Recombinant Human GNPNAT1 Protein: Functions & Applications

Recombinant Human GNPNAT1 Protein is a recombinantly produced form of glucosamine-phosphate N-acetyltransferase 1, a critical enzyme in the hexosamine biosynthesis pathway responsible for catalyzing the acetylation of glucosamine-6-phosphate to form N-acetylglucosamine-6-phosphate.

Protein Name

Glucosamine-Phosphate N-Acetyltransferase 1 (GNPNAT1)

Also Known As

GNA1; Glucosamine-6-phosphate acetyltransferase

Biological Function

Catalyzes acetylation of glucosamine-6-phosphate to N-acetylglucosamine-6-phosphate in the hexosamine biosynthesis pathway

Pathway Involvement

Hexosamine biosynthesis pathway; UDP-GlcNAc biosynthesis; O-GlcNAc glycosylation

Product SKU

rp371606

Species

Human (Homo sapiens)

Protein Type

Recombinant protein

What Is GNPNAT1? Gene, Protein, and Biological Role

GNPNAT1 (Glucosamine-Phosphate N-Acetyltransferase 1), also referred to as GNA1, encodes an enzyme that belongs to the acetyltransferase superfamily. The protein catalyzes a pivotal reaction in the hexosamine biosynthesis pathway (HBP): the transfer of an acetyl group from acetyl-CoA to glucosamine-6-phosphate (GlcN-6P), yielding N-acetylglucosamine-6-phosphate (GlcNAc-6P).

This reaction is an obligatory step in the de novo synthesis of uridine diphosphate N-acetylglucosamine (UDP-GlcNAc), a high-energy sugar nucleotide that serves as the essential donor substrate for O-GlcNAc transferase (OGT) and other glycosyltransferases. UDP-GlcNAc is required for the O-GlcNAcylation of nuclear and cytoplasmic proteins, a post-translational modification linked to nutrient sensing, cellular stress responses, and transcriptional regulation.

Because the HBP integrates inputs from glucose, glutamine, acetyl-CoA, and UTP metabolism, GNPNAT1 sits at a metabolic intersection of broad physiological relevance, including diabetes, cancer metabolism, and neurodegeneration research.

Biochemical Properties of Recombinant Human GNPNAT1 Protein

Recombinant Human GNPNAT1 Protein (SKU: rp371606) is produced using recombinant expression systems to ensure consistent quality and biological activity suitable for in vitro research applications. The protein corresponds to the human GNPNAT1 gene product and retains the acetyltransferase catalytic domain characteristic of the GNAT (GCN5-related N-acetyltransferase) superfamily.

Key biochemical characteristics include:

• Enzymatic activity: Glucosamine-6-phosphate acetyltransferase activity, requiring acetyl-CoA as co-substrate.
• Substrate specificity: Highly specific for glucosamine-6-phosphate; activity is dependent on the availability of acetyl-CoA.
• Expression system: Recombinant production enables high purity and lot-to-lot reproducibility. Specific expression host and purity data should be confirmed in the product datasheet.

Researchers are advised to consult the current product datasheet (available from Aladdin Scientific) for confirmed specifications including molecular weight, purity grade, formulation buffer, storage conditions, and recommended reconstitution protocols.

Research Applications of Recombinant Human GNPNAT1 Protein

Recombinant Human GNPNAT1 Protein is a versatile reagent suited to multiple research contexts where the hexosamine biosynthesis pathway, O-GlcNAc signaling, or UDP-GlcNAc metabolism is under investigation:

• Enzyme activity assays: Used as an active enzyme standard or positive control in colorimetric, fluorometric, or coupled enzymatic assays measuring acetyltransferase activity.
• Biochemical pathway reconstitution: Enables in vitro reconstruction of HBP steps to study flux, enzyme kinetics, and inhibitor screening.
• Drug discovery and inhibitor screening: Serves as a target enzyme for identifying small-molecule inhibitors relevant to metabolic diseases, cancer, and infection biology where HBP dysregulation is implicated.
• Protein interaction studies: Can be used in co-immunoprecipitation (Co-IP), pull-down assays, or surface plasmon resonance (SPR) to map protein–protein interactions within the HBP.
• Structural biology: May serve as a source protein for crystallography or cryo-EM studies aimed at resolving the GNPNAT1 active site for structure-based drug design.
• Antibody validation: Used as a positive control antigen when validating anti-GNPNAT1 antibodies by Western blot or ELISA.

Handling, Storage, and Reconstitution Guidelines

Proper handling and storage of Recombinant Human GNPNAT1 Protein are essential to maintaining enzymatic activity and structural integrity. General best practices for recombinant proteins apply:

• Storage: Store lyophilized protein at –20 °C or –80 °C as specified in the product datasheet. Avoid repeated freeze–thaw cycles, which can cause protein aggregation and loss of activity.
• Reconstitution: Reconstitute in the buffer recommended in the product datasheet (typically sterile PBS or manufacturer-specified buffer). Allow the vial to equilibrate to room temperature before opening. Centrifuge briefly before reconstitution to collect lyophilized material.
• Working aliquots: Prepare single-use aliquots immediately after reconstitution and store at –80 °C to maximize shelf life.
• Carrier protein: If the protein is sensitive to dilute concentrations, consider adding 0.1% BSA (carrier-protein grade) to stabilize the preparation.

Note: Always refer to the current Aladdin Scientific product datasheet for lot-specific storage conditions, formulation details, and reconstitution volumes.

Frequently Asked Questions About Recombinant Human GNPNAT1 Protein

See the FAQ section below for answers to common questions from researchers working with this product.

Frequently asked questions

What is the role of GNPNAT1 in the hexosamine biosynthesis pathway?

GNPNAT1 catalyzes the acetylation of glucosamine-6-phosphate to N-acetylglucosamine-6-phosphate using acetyl-CoA as the acetyl donor. This step is essential for the downstream synthesis of UDP-GlcNAc, the nucleotide sugar required for O-GlcNAcylation and other glycosylation events in the cell.

Which expression system is used to produce Recombinant Human GNPNAT1 Protein (rp371606)?

The specific expression host system for rp371606 should be confirmed by consulting the current Aladdin Scientific product datasheet, as expression systems (e.g., E. coli, baculovirus/insect cells, HEK293) can affect post-translational modifications and activity profiles. Please contact Aladdin Scientific technical support for the most up-to-date lot-specific information.

How should Recombinant Human GNPNAT1 Protein be stored to preserve activity?

As a general best practice for recombinant proteins, store the lyophilized product at –20 °C or –80 °C and avoid repeated freeze–thaw cycles. After reconstitution, prepare single-use aliquots and store at –80 °C. Always consult the product-specific datasheet for confirmed storage and stability conditions.

Can Recombinant Human GNPNAT1 Protein be used for inhibitor screening assays?

Yes, Recombinant Human GNPNAT1 Protein is suitable for use as a target enzyme in inhibitor screening assays. Researchers can employ coupled enzymatic or direct acetyltransferase activity assays to measure inhibition of GlcN-6P acetylation. Such screens are relevant to drug discovery programs targeting metabolic diseases and cancer where HBP flux is dysregulated.

Is Recombinant Human GNPNAT1 Protein suitable for use as a Western blot positive control?

Yes, the recombinant protein can be used as a positive control antigen to validate anti-GNPNAT1 antibodies by Western blot or ELISA, provided the molecular weight and tag configuration are compatible with the detection system being used. Verify the protein's tag and formulation in the product datasheet before use.

References

1. Boehmelt G, et al. Decreased UDP-GlcNAc levels abrogate proliferation control in EMeg32-deficient cells. EMBO J. 2000.
2. Vosseller K, et al. Elevated nucleocytoplasmic glycosylation by O-GlcNAc results in insulin resistance associated with defects in Akt activation in 3T3-L1 adipocytes. Proc Natl Acad Sci USA. 2002.
3. Hart GW, Slawson C, Ramirez-Correa G, Lagerlof O. Cross talk between O-GlcNAcylation and phosphorylation: roles in signaling, transcription, and chronic disease. Annu Rev Biochem. 2011.

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