Recombinant Human EFNA3 protein(Met1-Ser213), His-tagged

• Cat.No.: EFNA3-3151H
• Product Overview: Recombinant Human EphrinA3 (NP_004943.1) (Met 1-Ser 213) with the C-terminal propeptide removed was expressed in HEK293, with a polyhistidine tag at the C-terminus.

Specification

• Species: Human
• Source: HEK293
• Tag: His
• Protein Length: 1-213 a.a.
• Form: Lyophilized from sterile PBS, pH 7.4. Normally 5 % - 8 % trehalose, mannitol and 0.01% Tween80 are added as protectants before lyophilization.
• Bio-activity: Measured by its ability to compete with human EphrinA3 / Fc for binding to immobilized mouse EphA6-his in a functional ELISA assay.
• Molecular Mass: The recombinant human EphrinA3 consists of 202 amino acids after removal of the signal peptide and has a predicted molecular mass of 23 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhEphrinA3 is approximately 35-40 kDa due to glycosylation.
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method
• Purity: > 95 % as determined by SDS-PAGE
• Storage: Samples are stable for up to twelve months from date of receipt at -20°C to -80°C. Store it under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
• Reconstitution: It is recommended that sterile water be added to the vial to prepare a stock solution of 0.2 ug/ul. Centrifuge the vial at 4°C before opening to recover the entire contents.

Gene Information

• Gene Name: EFNA3 ephrin-A3 [ Homo sapiens ]
• Official Symbol: EFNA3
• Synonyms: EFNA3; ephrin-A3; EPLG3; Ehk1 L; LERK3; EFL-2; LERK-3; EHK1 ligand; ligand of eph-related kinase 3; eph-related receptor tyrosine kinase ligand 3; EFL2; Ehk1-L
• Gene ID: 1944
• mRNA Refseq: NM_004952
• Protein Refseq: NP_004943
• MIM: 601381
• UniProt ID: P52797

Case Study

Case 1: Yiminniyaze R, et al. Cancer Med. 2023

EphrinA3 drives lung adenocarcinoma progression via EphA1/A10 receptors, activating AKT/ERK/p38MAPK pathways to promote EMT and MMP-2/-9 upregulation. High EphrinA3 expression correlates with poor LUAD prognosis, enhancing tumor growth and metastasis in vitro and in vivo, positioning it as a therapeutic target for LUAD invasion and survival outcomes.

Fig1. EphrinA3 protein expression levels in BEAS‐2B and LUAD cells.

Fig2. EphrinA3 promotes CyclinD1 expression.

Case 2: Singal CMS, et al. ASN Neuro. 2021

HIV-1 Tat upregulates astrocytic ephrinA3, impairing EAAT1/2-mediated glutamate uptake and exacerbating neurotoxicity in HIV-associated neurodegeneration. EphrinA3 knockdown restores glutamate clearance and neuronal survival, validated in HIV-infected patient brains, identifying the ephrinA3-EphA4 axis as a therapeutic target for neurocognitive disorders and excitotoxicity management.

Fig1. Expression of ephrinA3 is upregulated when transfected with HIV-1 Tat B.

Fig2. The blots were probed with ephrinA3 and β-tubulin antibody and significant knockdown was observed at 40 pmol concentration as compared to that of scrambled control.

Application

1. Therapeutic Potential of Recombinant EFNA3 Protein in Neuroprotection and Oncology Recombinant ephrin-A3 (EFNA3) protein emerges as a dual-targeted therapy for neurocognitive disorders and cancer, modulating Eph receptor signaling to address glutamate excitotoxicity in HIV-associated neurodegeneration or tumor progression. In neuroHIV models, EFNA3 normalization restores astrocytic EAAT1/2 function, reducing extracellular glutamate and neuronal death, while in oncology, it may suppress EphA4-driven metastasis by regulating cell-cell adhesion and invasion pathways. 2. Mechanistic Insights and Preclinical Validation Studies demonstrate EFNA3’s role in reversing HIV Tat-induced glutamate transporter downregulation via EphA4 receptor interaction, mitigating excitotoxicity in human astrocyte-neuron co-cultures. In glioblastoma and lung adenocarcinoma, EFNA3 overexpression inhibits tumor migration by destabilizing focal adhesions and MMP activity. Preclinical models highlight its dose-dependent efficacy, balancing neuroprotection with anti-metastatic effects without disrupting physiological Eph signaling. 3. Delivery Challenges and Combinatorial Strategies Current limitations include poor blood-brain barrier penetration and transient activity. Innovations like engineered exosomes or CRISPR-activated EFNA3 expression systems aim to enhance CNS delivery and sustained action. Pairing recombinant EFNA3 with antiretroviral therapies or immune checkpoint inhibitors could synergize outcomes in neuroHIV or resistant cancers, advancing precision medicine for complex pathologies.

Fig1. Glutamate uptake is regulated by ephrinA3 by maintaining the levels of glutamate transporters (EAAT1 and EAAT2) in normal brain cells. (Chitra Mohinder Singh Singal, 2021)