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GRK5 cell lysate

Human GRK5 Baculovirus-Insect cells Overexpression Lysate

Gene Names
GRK5; GPRK5
Applications
Western Blot
Synonyms
GRK5; Human GRK5 Baculovirus-Insect cells Overexpression Lysate; Human GRK5/GPRK5 Insect Cell Lysate (WB positive control); G protein-coupled receptor kinase 5; Human GPRK5 Overexpression Lysate; GRK5 cell lysate
Ordering
For Research Use Only!
Host
Baculovirus-Insect cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
600aa
Sequence
Met1-Ser590
Applicable Applications for GRK5 cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the human GRK5 (NP_005299.1) (Met 1-Ser 590) was fused with a polyhistidine tag at the C-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for GRK5 cell lysate
G protein-coupled receptor kinase 5, also known as G protein-coupled receptor kinase GRK5 and GRK5, is a member of the protein kinase superfamily, AGC Ser/Thr protein kinase family, and GPRK subfamily. GRKs specifically phosphorylate agonist-occupied G protein-coupled receptors at the inner surface of the plasma membrane (PM), leading to receptor desensitization. GRKs utilize a variety of mechanisms to bind tightly, and sometimes reversibly, to cellular membranes. GRKs play an important role in mediating agonist-specific desensitization of numerous G protein-coupled receptors.
GRK5 contains one AGC-kinase C-terminal domain, one protein kinase domain, and one RGS domain. GRK5 specifically phosphorylates the activated forms of G protein-coupled receptors. Phospholipid-stimulated autophosphorylation may represent a novel mechanism for membrane association and regulation of GRK5 activity. GRK5 deficiency significantly exaggerates microgliosis and astrogliosis in the presence of an inflammatory initiator, such as the excess fibrillar Abeta and the subsequent active inflammatory reactions. GRK5 deficiency has been linked to early Alzheimer's disease in humans and mouse models of the disease.

This Human GRK5 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of GRK5 protein from the overexpression lysate was verified.
References
Kunapuli, P. et al., 1994, J Biol Chem. 269 (14):10209-12. Millman, E.E. et al., 2004, Br J Pharmacol 141 (2):277-84. Thiyagarajan, M.M. et al., 2004, J Biol Chem 279 (17):17989-95. Suo, Z. et al., 2007, Neurobiol Aging. 28 (12):1873-88. Li, L. et al., 2008, J Neuroinflammation. 5 :24.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
67,787 Da
NCBI Official Full Name
G protein-coupled receptor kinase 5
NCBI Official Synonym Full Names
G protein-coupled receptor kinase 5
NCBI Official Symbol
GRK5
NCBI Official Synonym Symbols
GPRK5
NCBI Protein Information
G protein-coupled receptor kinase 5; FP2025; g protein-coupled receptor kinase GRK5
UniProt Protein Name
G protein-coupled receptor kinase 5
UniProt Gene Name
GRK5
UniProt Synonym Gene Names
GPRK5
UniProt Entry Name
GRK5_HUMAN

NCBI Description

This gene encodes a member of the guanine nucleotide-binding protein (G protein)-coupled receptor kinase subfamily of the Ser/Thr protein kinase family. The protein phosphorylates the activated forms of G protein-coupled receptors thus initiating their deactivation. It has also been shown to play a role in regulating the motility of polymorphonuclear leukocytes (PMNs). [provided by RefSeq, Jul 2008]

Uniprot Description

GRK5: Serine/threonine kinase that phosphorylates preferentially the activated forms of a variety of G-protein- coupled receptors (GPCRs). Such receptor phosphorylation initiates beta-arrestin-mediated receptor desensitization, internalization, and signaling events leading to their down-regulation. Phosphorylates a variety of GPCRs, including adrenergic receptors, muscarinic acetylcholine receptors (more specifically Gi-coupled M2/M4 subtypes), dopamine receptors and opioid receptors. In addition to GPCRs, also phosphorylates various substrates: Hsc70- interacting protein/ST13, TP53/p53, HDAC5, and arrestin-1/ARRB1. Phosphorylation of ARRB1 by GRK5 inhibits G-protein independent MAPK1/MAPK3 signaling downstream of 5HT4-receptors. Phosphorylation of HDAC5, a repressor of myocyte enhancer factor 2 (MEF2) leading to nuclear export of HDAC5 and allowing MEF2- mediated transcription. Phosphorylation of TP53/p53, a crucial tumor suppressor, inhibits TP53/p53-mediated apoptosis. Phosphorylation of ST13 regulates internalization of the chemokine receptor. Phosphorylates rhodopsin (RHO) (in vitro) and a non G- protein-coupled receptor, LRP6 during Wnt signaling (in vitro). Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. GPRK subfamily.

Protein type: Protein kinase, Ser/Thr (non-receptor); Protein kinase, AGC; EC 2.7.11.16; Kinase, protein; AGC group; GRK family; GRK subfamily

Chromosomal Location of Human Ortholog: 10q26.11

Cellular Component: nucleoplasm; nuclear membrane; focal adhesion; cytoplasm; plasma membrane; nucleus

Molecular Function: G-protein coupled receptor kinase activity; protein serine/threonine kinase activity; protein binding; protein kinase C binding; phospholipid binding; ATP binding

Biological Process: G-protein signaling, coupled to cAMP nucleotide second messenger; tachykinin signaling pathway; Wnt receptor signaling pathway; apoptosis; protein amino acid autophosphorylation; regulation of G-protein coupled receptor protein signaling pathway; negative regulation of apoptosis

Research Articles on GRK5

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Product Notes

The GRK5 grk5 (Catalog #AAA8114177) is a Cell Lysate produced from Baculovirus-Insect cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 600aa. AAA Biotech's GRK5 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the GRK5 grk5 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Ser59 0. It is sometimes possible for the material contained within the vial of "GRK5, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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