The catalytic subunit of protein phosphatase 2A (PP2A) is inactivated by in vitro phosphorylation of Tyr-307 by receptor and nonreceptor protein tyrosine kinases. The catalytic subunit of PP2A is phosphorylated by tyrosine-specific protein kinases and associates with a variety of regulatory subunits. Phosphorylation is enhanced in the presence of the phosphatase inhibitor okadaic acid, consistent with an autodephosphorylation reaction. Phosphorylation is catalyzed by p60v-src, p56lck, epidermal growth factor receptors, and insulin receptors. Transient deactivation of PP2A might enhance transmission of cellular signals through kinase cascades within cells. In eukaryotes, the phosphorylation and dephosphorylation of proteins on serine and threonine residues is an essential means of regulating a broad range of cellular functions, including cell division, homeostasis and apoptosis. A group of proteins that are intimately involved in this process are the protein phosphatases.
Background References
1. Lei X et al. PP2A and tumor radiotherapy. Hereditas. 2020 Aug
2. Ronk H et al. Targeting PP2A for cancer therapeutic modulation. Cancer Biol Med. 2022 Nov
Sequence Similarity
Belongs to the PPP phosphatase family. PP-1 subfamily.
Post-translational Modification
Reversibly methyl esterified on Leu-309 by leucine carboxyl methyltransferase 1 (LCMT1) and protein phosphatase methylesterase 1 (PPME1). Carboxyl methylation influences the affinity of the catalytic subunit for the different regulatory subunits, thereby modulating the PP2A holoenzyme's substrate specificity, enzyme activity and cellular localization.; Phosphorylation of either threonine (by autophosphorylation-activated protein kinase) or tyrosine results in inactivation of the phosphatase. Auto-dephosphorylation has been suggested as a mechanism for reactivation.; May be monoubiquitinated by NOSIP.
Subcellular Location
Cytoplasm, Nucleus, Chromosome, centromere, cytoskeleton, spindle pole.
Western blot analysis of Phospho-PP2A (Y307) on different lysates with Rabbit anti-Phospho-PP2A (Y307) antibody (ET1609-40) at 1/1,000 dilution.
Lane 1: A431 (Human epidermoid carcinoma skin squamous cell) cell lysate Lane 2: F9 (Mouse teratocarcinoma cell) cell lysate
Lysates/proteins at 10 µg/Lane. Exposure time: 30 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature Primary antibody: ET1609-40, 1/1,000 in 5% NFDM/TBST, overnight at 4 ℃ Secondary antibody: Goat Anti-Rabbit IgG-HRP(HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 35.6 kDa Observed band size: 36 kDa
ICC staining of Phospho-PP2A (Y307) in PC-12 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 (ET1609-40, 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 human pancreas tissue using anti-Phospho-PP2A (Y307) 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 (ET1609-40, 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 mouse brain tissue using anti-Phospho-PP2A (Y307) 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 (ET1609-40, 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 mouse pancreas tissue using anti-Phospho-PP2A (Y307) 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 (ET1609-40, 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 kidney tissue with Rabbit anti-Phospho-PP2A (Y307) antibody (ET1609-40) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-40) at 1/200 dilution for 1 hour 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 esophagus tissue with Rabbit anti-Phospho-PP2A (Y307) antibody (ET1609-40) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1609-40) at 1/200 dilution for 1 hour 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.
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