Recombinant Human GBA2 Protein: Research Use Guideline

Recombinant Human GBA2 Protein is a non-lysosomal glucosylceramidase used in research to investigate sphingolipid metabolism, neurological disease mechanisms, and glucosylceramide hydrolysis pathways.

Product Name

Recombinant Human GBA2 Protein

SKU

rp372119

Protein Function

Non-lysosomal glucosylceramidase that hydrolyzes glucosylceramide to ceramide and glucose at the cell surface and endoplasmic reticulum

Research Relevance

Implicated in sphingolipid homeostasis, male fertility, neurodegeneration, and hereditary spastic paraplegia

Species

Human (Homo sapiens)

Recommended Storage

Store at -80°C upon receipt; avoid repeated freeze-thaw cycles

Intended Use

For research use only; not for therapeutic or diagnostic use

Protein Background and Biological Significance

GBA2 (non-lysosomal glucosylceramidase 2), encoded by the GBA2 gene, is a membrane-associated enzyme that catalyzes the hydrolysis of glucosylceramide (GlcCer) into ceramide and glucose. Unlike the lysosomal glucocerebrosidase GBA1, GBA2 is localized primarily at the endoplasmic reticulum and plasma membrane, functioning at neutral to slightly acidic pH conditions. This spatial and pH distinction makes GBA2 a critical regulator of extracellular and non-lysosomal sphingolipid pools.

Dysregulation of GBA2 activity has been linked to multiple disease contexts. Loss-of-function mutations in GBA2 are associated with hereditary spastic paraplegia (SPG46) and cerebellar ataxia. GBA2 has also been implicated in male fertility, as it contributes to the regulation of glucosylceramide levels in spermatocytes and is necessary for normal sperm morphology. Furthermore, there is growing interest in the relationship between GBA2 activity and Parkinson's disease pathology, where imbalances in ceramide and glucosylceramide metabolism may influence alpha-synuclein aggregation. Recombinant Human GBA2 Protein provides researchers with a purified, well-characterized tool to investigate these disease-relevant biochemical pathways in vitro.

Protein Handling, Reconstitution, and Storage

Proper handling of Recombinant Human GBA2 Protein is essential to preserve enzymatic activity and ensure experimental reproducibility. Upon receipt, the protein should be briefly centrifuged to consolidate any material that may have adhered to the tube walls during shipping. If supplied in lyophilized form, reconstitute gently using the buffer recommended in the product datasheet (typically sterile PBS or Tris-based buffer); allow the protein to dissolve slowly at 4°C with gentle swirling and avoid vigorous vortexing, which can cause protein denaturation.

For short-term storage (up to one week), the reconstituted protein may be kept at 4°C. For long-term storage, aliquot into single-use volumes to minimize freeze-thaw cycles and store at -80°C. Repeated freeze-thaw cycles significantly reduce enzymatic activity and should be strictly avoided. Adding a carrier protein such as 0.1% BSA to the reconstitution buffer can help stabilize dilute preparations. Always work on ice when setting up enzymatic assays to minimize activity loss prior to use.

Note: Specific reconstitution volumes, buffer compositions, and concentration data are provided in the lot-specific Certificate of Analysis (CoA). Researchers should consult the CoA for exact specifications of each received lot.

Experimental Applications and Assay Guidance

Recombinant Human GBA2 Protein is suitable for a range of in vitro research applications centered on sphingolipid biology and enzyme biochemistry:

• Enzymatic activity assays: GBA2 activity can be measured fluorometrically using artificial substrates such as 4-methylumbelliferyl-β-D-glucopyranoside (4-MUG) or mass spectrometry-based methods with natural substrate glucosylceramide. Inclusion of conduritol B epoxide (CBE) in parallel reactions is recommended to selectively inhibit GBA1 activity, thereby isolating GBA2-specific activity in mixed systems.
• Inhibitor screening: The recombinant protein is well-suited for testing novel small-molecule inhibitors of GBA2 in drug discovery contexts, particularly for conditions such as hereditary spastic paraplegia and Niemann-Pick type C disease.
• Protein–protein interaction studies: Pull-down and co-immunoprecipitation experiments can be designed to identify GBA2 binding partners relevant to sphingolipid trafficking.
• Western blot and ELISA standards: Recombinant GBA2 protein can serve as a positive control or calibration standard in immunodetection assays.
• Structural and biophysical studies: The purified protein supports circular dichroism (CD), thermal shift assays, and other biophysical characterization experiments.

Optimal assay conditions (pH, temperature, substrate concentrations, and buffer composition) should be empirically determined or referenced from validated published protocols, as conditions may vary by experimental design.

Quality, Safety, and Regulatory Considerations

Quality: Recombinant Human GBA2 Protein (rp372119) is produced under controlled conditions with quality assessed by SDS-PAGE purity analysis and functional activity testing. Researchers should request the Certificate of Analysis (CoA) for each lot to confirm purity, concentration, and specific activity data before initiating experiments. Lot-to-lot consistency should be evaluated when transitioning between lots in ongoing studies.

Safety: This protein is intended for in vitro research use only. Standard laboratory safety practices should be observed when handling recombinant proteins, including the use of appropriate personal protective equipment (PPE) such as gloves, lab coat, and eye protection. Although recombinant GBA2 is not classified as a hazardous material under standard laboratory regulations, all biological materials should be treated with appropriate caution. Refer to the Safety Data Sheet (SDS) provided by Aladdin Scientific for specific hazard information.

Regulatory: This product is for research use only (RUO) and is not approved for use in clinical diagnostics, therapeutic applications, or administration to humans or animals. Researchers are responsible for ensuring compliance with all applicable institutional biosafety protocols and local regulatory requirements governing the use of recombinant proteins in their facility.

Frequently asked questions

What is the difference between GBA1 and GBA2?

GBA1 (acid beta-glucosidase) is a lysosomal enzyme that hydrolyzes glucosylceramide within the lysosome and is deficient in Gaucher's disease. GBA2 is a non-lysosomal glucosylceramidase localized at the endoplasmic reticulum and plasma membrane, functioning at neutral pH. They are structurally distinct enzymes encoded by different genes with different inhibitor sensitivities; notably, GBA2 is inhibited by N-butyldeoxynojirimycin (NB-DNJ) but not by conduritol B epoxide (CBE), whereas GBA1 is inhibited by CBE.

How should I verify enzymatic activity of the recombinant GBA2 protein after reconstitution?

A standard fluorometric assay using 4-methylumbelliferyl-β-D-glucopyranoside (4-MUG) as an artificial substrate is commonly used to confirm GBA2 activity. Reactions should be conducted at the pH optimum appropriate for GBA2 (typically near neutral pH) and compared with a no-enzyme blank. To specifically attribute activity to GBA2, include conduritol B epoxide (CBE) to inhibit any GBA1 contamination. Consult the lot-specific CoA for activity specifications.

Can Recombinant Human GBA2 Protein be used for in vivo studies?

No. Recombinant Human GBA2 Protein (rp372119) is designated for research use only (RUO) and is not validated, approved, or intended for in vivo administration to humans or animals, nor for use in clinical diagnostics or therapeutic applications. All use of this product should conform to applicable institutional biosafety and regulatory guidelines.

How many freeze-thaw cycles can the recombinant GBA2 protein tolerate?

It is strongly recommended to minimize freeze-thaw cycles to preserve enzymatic activity. Best practice is to aliquot the protein into single-use volumes upon first thaw and store aliquots at -80°C. As a general guideline for recombinant enzymes, more than two to three freeze-thaw cycles can lead to significant activity loss; however, the exact tolerance for GBA2 should be confirmed from the product-specific CoA or by empirical testing in your laboratory.

What diseases are associated with GBA2 and why is it a research target?

Mutations in GBA2 are associated with hereditary spastic paraplegia type 46 (SPG46), autosomal recessive cerebellar ataxia, and male infertility due to impaired sperm morphology. GBA2 has also attracted interest in the context of Parkinson's disease research due to its role in glucosylceramide and ceramide homeostasis, which may intersect with alpha-synuclein biology. Additionally, GBA2 inhibition has been explored as a therapeutic strategy in lysosomal storage disorders such as Gaucher disease and Niemann-Pick type C disease.

References

1. Körschen HG, Yildiz Y, Raju DN, et al. The non-lysosomal beta-glucosidase GBA2 is a non-integral membrane-associated protein at the endoplasmic reticulum (ER) and Golgi. J Biol Chem. 2013;288(5):3381-3393. doi:10.1074/jbc.M112.414714
2. Boot RG, Verhoek M, Donker-Koopman W, et al. Identification of the non-lysosomal glucosylceramidase as beta-glucosidase 2. J Biol Chem. 2007;282(2):1305-1312. doi:10.1074/jbc.M610544200
3. Martin E, Schüle R, Smets K, et al. Loss of function of glucocerebrosidase GBA2 is responsible for motor neuron defects in hereditary spastic paraplegia. Am J Hum Genet. 2013;92(2):238-244. doi:10.1016/j.ajhg.2012.11.021
4. Yildiz Y, Matern H, Thompson B, et al. Mutation of beta-glucosidase 2 causes glycolipid storage disease and impaired male fertility. J Clin Invest. 2006;116(11):2985-2994. doi:10.1172/JCI29224

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