The glyoxal pathway plays a role in the detoxification of glucose degradation products (GDP). Glyoxalase I (GLO1), a member of the glyoxalase family, is effective in eliminating GDP. Overexpression or silencing of Glyoxalase I in mice brain suggests an association between Glyoxalase I and anxiety. Glyoxalase I has three isoforms generated from two alleles in the genome which forms two homodimers and one heterodimer, each subunit binding one zinc ion. Research demonstrates that GLO1 gene expression is induced in colon carcinoma. Both an insulin response element (IRE), and a zinc metal response element (MRE) in the promoter region of the GLO1 gene have been identified.
Background References
1. de Hemptinne V et al. Phosphorylation on Thr-106 and NO-modification of glyoxalase I suppress the TNF-induced transcriptional activity of NF-kappaB. Mol Cell Biochem 325:169-178 (2009).
2. Ridderstroem M et al. Involvement of an active-site Zn2+ ligand in the catalytic mechanism of human glyoxalase I. J Biol Chem 273:21623-21628 (1998).
Sequence Similarity
Belongs to the glyoxalase I family.
Post-translational Modification
Glutathionylation at Cys-139 inhibits enzyme activity.; Phosphorylated at Thr-107 in the presence of CaMK2. However, this is a consensus site for phosphorylation by CK2 so phosphorylation may be mediated by CK2 rather than CaMK2. Phosphorylation is induced by TNF and suppresses the TNF-induced transcriptional activity of NF-kappa-B.; Exists in a nitric oxide (NO)-modified form. The exact nature of the modification is unknown, but it suppresses the TNF-induced transcriptional activity of NF-kappa-B.
Western blot analysis of GLO1 on different lysates with Rabbit anti-GLO1 antibody (ET7106-68) at 1/1,000 dilution.
Lane 1: 293T cell lysate (20 µg/Lane) Lane 2: C2C12 cell lysate (20 µg/Lane) Lane 3: Mouse testis tissue lysate (40 µg/Lane) Lane 4: Rat spleen tissue lysate (40 µg/Lane)
Predicted band size: 21 kDa Observed band size: 21 kDa
Exposure time: 1 minute 2 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7106-68) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
☑ Knockdown (KD)
Western blot analysis of GLO1 on different lysates with Rabbit anti-GLO1 antibody (ET7106-68) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate Lane 2: HAP1-GLO1 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 21 kDa Observed band size: 21 kDa
Exposure time: 180 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET7106-68) at 1/1,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ICC staining of GLO1 in 293T cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-68, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of GLO1 in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-68, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ICC staining of GLO1 in SH-SY5Y cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET7106-68, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
Immunohistochemical analysis of paraffin-embedded rat epididymis tissue using anti-GLO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-68, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Immunohistochemical analysis of paraffin-embedded human thyroid tissue using anti-GLO1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET7106-68, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Flow cytometric analysis of GLO1 was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET7106-68, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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