Phosphatidyl-4-phosphate-5-kinase, gamma (PIP5K1g, PIP5K-Ig, IP5K1-gamma, PtdIns(4)P-5-kinase 1 gamma, EC 2.7.1.68)

Catalyzes the phosphorylation of phosphatidylinositol 4-phosphate (PtdIns4P) to form phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2). PtdIns(4,5)P2 is involved in a variety of cellular processes and is the substrate to form phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P3), another second messenger. The majority of PtdIns(4,5)P2 is thought to occur via type I phosphatidylinositol 4-phosphate 5-kinases given the abundance of PtdIns4P. Participates in a variety of cellular processes such as vesicle mediated transport, cell adhesion, cell polarization and cell migration. Together with PIP5K1A is required for phagocytosis, but they regulate different types of actin remodeling at sequential steps. Promotes particle attachment by generating the pool of PtdIns(4,5)P2 that induces controlled actin depolymerization to facilitate Fc-gamma-R clustering. Mediates RAC1-dependent reorganization of actin filaments. Required for synaptic vesicle transport. Controls the plasma membrane pool of PtdIns(4,5)P2 implicated in synaptic vesicle endocytosis and exocytosis. Plays a role in endocytosis mediated by clathrin and AP-2 (adaptor protein complex 2). Required for clathrin-coated pits assembly at the synapse. Participates in cell junction assembly. Modulates adherens junctions formation by facilitating CDH1 trafficking. Required for focal adhesion dynamics. Modulates the targeting of talins (TLN1 and TLN2) to the plasma membrane and their efficient assembly into focal adhesions. Regulates the interaction between talins (TLN1 and TLN2) and beta-integrins. Required for uropodium formation and retraction of the cell rear during directed migration. Has a role in growth factor- stimulated directional cell migration and adhesion. Required for talin assembly into nascent adhesions forming at the leading edge toward the direction of the growth factor. Negative regulator of T-cell activation and adhesion. Negatively regulates integrin alpha-L/beta-2 (LFA-1) polarization and adhesion induced by T-cell receptor. Together with PIP5K1A have a role during embryogenesis and together with PIP5K1B may have a role immediately after birth.\n\nApplication(s): \nSuitable for use in Western Blot, ELISA.\n\nRecommended Dilution:\nELISA: 1:50,000. Antibody titers and specificity were analyzed by ELISA assay using control samples (varying concentration of GST-EPOr) and control anti GST antibody. Affinity purified antibody was found to detect EPOr at concentrations of 50ng/ml. Neutralization of ligand binding has been detected using Erytholukemia cell line TF-1 (3000 EPO receptors/cell) to block Iodine-labeled erythropoietin ligand binding. \n\nWestern Blot: ~10ug/ml. Recognizes EPOr-GST at its migration of 49kD on SDS-PAGE. Good secondary anti-sheep reagents are essential for this assay. Recommended use of rabbit anti-sheep rather than donkey anti-sheep antibodies as a secondary labeling reagents.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

P4071-76

Size

100ug

Applications

ELISA, WB

Hosts

Rabbit

Reactivities

Hum

Form

Supplied as a liquid in PBS, pH 7.2, 1mg/ml BSA, 0.1% sodium azide before the addition of 40% glycerol.

P Type

Pab

Purity

Purified by immunoaffinity chromatography.

Isotype

IgG

References

1. Hiasmitsu I etal (1998) Type 1 Phosphatidylinositol-4- phosphate 5-Kinases. J Biol. Chem. 273 (15) 8741-8748\nGeneral References:\nhttp://www.uniprot.org/uniprot/O60331\n[1] \"Two novel phosphatidylinositol-4-phosphate 5-kinase type Igamma splice variants expressed in human cells display distinctive cellular targeting.\"\nSchill N.J., Anderson R.A.\nBiochem. J. 422:473-482(2009) [PubMed: 19548880] [Abstract]\nCited for: NUCLEOTIDE SEQUENCE [MRNA] (ISOFORMS 2 AND 3), SUBCELLULAR LOCATION, TISSUE SPECIFICITY. \n[5] \"Recruitment and regulation of phosphatidylinositol phosphate kinase type 1 gamma by the FERM domain of talin.\"\nDi Paolo G., Pellegrini L., Letinic K., Cestra G., Zoncu R., Voronov S., Chang S., Guo J., Wenk M.R., De Camilli P.\nNature 420:85-89(2002) [PubMed: 12422219] [Abstract]\nCited for: FUNCTION, INTERACTION WITH TLN2, SUBCELLULAR LOCATION, ENZYME REGULATION. \n[6] \"ARF6 stimulates clathrin/AP-2 recruitment to synaptic membranes by activating phosphatidylinositol phosphate kinase type Igamma.\"\nKrauss M., Kinuta M., Wenk M.R., De Camilli P., Takei K., Haucke V.\nJ. Cell Biol. 162:113-124(2003) [PubMed: 12847086] [Abstract]\nCited for: FUNCTION, INTERACTION WITH ARF6, SUBCELLULAR LOCATION. \n[7] \"Regulation of the interaction between PIPKI gamma and talin by proline-directed protein kinases.\"\nLee S.Y., Voronov S., Letinic K., Nairn A.C., Di Paolo G., De Camilli P.\nJ. Cell Biol. 168:789-799(2005) [PubMed: 15738269] [Abstract]\nCited for: PHOSPHORYLATION AT TYR-649 AND SER-650, MUTAGENESIS OF SER-650. \n[8] \"Type I gamma phosphatidylinositol phosphate kinase modulates adherens junction and E-cadherin trafficking via a direct interaction with mu 1B adaptin.\"\nLing K., Bairstow S.F., Carbonara C., Turbin D.A., Huntsman D.G., Anderson R.A.\nJ. Cell Biol. 176:343-353(2007) [PubMed: 17261850] [Abstract]\nCited for: FUNCTION, SUBCELLULAR LOCATION, INTERACTION WITH CDH1. \n[9] \"Type I gamma phosphatidylinositol phosphate kinase is required for EGF-stimulated directional cell migration.\"\nSun Y., Ling K., Wagoner M.P., Anderson R.A.\nJ. Cell Biol. 178:297-308(2007) [PubMed: 17635937] [Abstract]\nCited for: FUNCTION IN CELL MIGRATION AND ADHESION. \n[10] \"SIRT1 regulates thyroid-stimulating hormone release by enhancing PIP5Kgamma activity through deacetylation of specific lysine residues in mammals.\"\nAkieda-Asai S., Zaima N., Ikegami K., Kahyo T., Yao I., Hatanaka T., Iemura S., Sugiyama R., Yokozeki T., Eishi Y., Koike M., Ikeda K., Chiba T., Yamaza H., Shimokawa I., Song S.Y., Matsuno A., Mizutani A. Setou M.\nPLoS ONE 5:E11755-E11755(2010) [PubMed: 20668706] [Abstract]\nCited for: ACETYLATION AT LYS-265 AND LYS-268, DEACETYLATION BY SIRT1. \n[11] \"PIP5K-driven PtdIns(4,5)P2 synthesis: regulation and cellular functions.\"\nvan den Bout I., Divecha N.\nJ. Cell Sci. 122:3837-3850(2009) [PubMed: 19889969] [Abstract]\nCited for: REVIEW ON FUNCTION. \n[12] \"Lethal contractural syndrome type 3 (LCCS3) is caused by a mutation in PIP5K1C, which encodes PIPKI gamma of the phophatidylinsitol pathway.\"\nNarkis G., Ofir R., Landau D., Manor E., Volokita M., Hershkowitz R., Elbedour K., Birk O.S.\nAm. J. Hum. Genet. 81:530-539(2007) [PubMed: 17701898] [Abstract]\nCited for: VARIANT LCCS3 ASN-253, CHARACTERIZATION OF VARIANT LCCS3 ASN-253.

Additional Info

Binds to a 85-90kD human protein. Binding can be blocked by the peptide immunogen.