G protein-coupled Receptor Kinase 6 (GRK6) is one of 7 members of the GRK serine/threonine kinase subfamily, which are known primarily for their role in desensitizing activated G protein-coupled receptors (GPCRs) . GRKs function by phosphorylating serine/threonine residues in activated GPCRs. Upon phosphorylation these residues serve as binding sites for β-arrestin proteins, inhibiting re-activation of GPCRs by blocking their re-association with G proteins . There is evidence that GRKs can also modulate selected non-GPCR signaling pathways . For example, GRK6 has been shown to modulate the Wnt signaling pathway via phosphorylation of LRP6 , and the insulin-like growth factor signaling pathway. GRK6 may also play a role in immune system function. Investigators have found GRK6 expression is typically abundant in hematopoietic tumor cell lines, and a recent research study demonstrated that GRK6 suppression was selectively lethal for a number of myeloma tumor cell lines .
Background References
1. Mushegian A, Gurevich VV, Gurevich EV. The origin and evolution of G protein-coupled receptor kinases. PLoS One. 2012;7(3):e33806. doi: 10.1371/journal.pone.0033806. Epub 2012 Mar 19.
2. Gurevich EV, Tesmer JJ, Mushegian A, Gurevich VV. G protein-coupled receptor kinases: more than just kinases and not only for GPCRs. Pharmacol Ther. 2012 Jan;133(1):40-69.
3. Zheng H, Worrall C, Shen H, Issad T, Seregard S, Girnita A, Girnita L. Selective recruitment of G protein-coupled receptor kinases (GRKs) controls signaling of the insulin-like growth factor 1 receptor. Proc Natl Acad Sci U S A. 2012 May 1;109(18):7055-60.
Western blot analysis of GRK6 on different lysates with Rabbit anti-GRK6 antibody (HA721884) at 1/1,000 dilution.
Lane 1: A549-si NT cell lysate Lane 2: A549-si GRK6 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 66 kDa Observed band size: 66 kDa
Exposure time: 3 minutes; 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 (HA721884) 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.
Western blot analysis of GRK6 on different lysates with Rabbit anti-GRK6 antibody (HA721884) at 1/1,000 dilution.
Lane 1: HeLa cell lysate Lane 2: 293T cell lysate Lane 3: Jurkat cell lysate Lane 4: MCF7 cell lysate Lane 5: K-562 cell lysate Lane 6: A549 cell lysate
Lysates/proteins at 30 µg/Lane.
Predicted band size: 66 kDa Observed band size: 66 kDa
Exposure time: 5 minutes;
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 (HA721884) 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.
Flow cytometric analysis of 293T cells labeling GRK6.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA721884, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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