Involvement of the C-terminal Proline-rich Motif of G Protein-coupled Receptor Kinases in Recognition of Activated Rhodopsin

Gan, Xiaoqing; Ma, Zhuguo; Deng, Ning; Wang, Jiyong; Ding, Jianping; Li, Lin

doi:10.1074/jbc.m407570200

Cited by 23 publications

(21 citation statements)

References 36 publications

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“…The sequence has one more distinguished region, the threonine/proline-rich motif, that is found also in other metazoan receptors (34). In the present study, the LPS-interacting protein was shown to bind LPS in a specific manner; no effect of the recombinant protein could be seen on cell-cell and celltissue adhesion.…”

Section: Discussionmentioning

confidence: 60%

Innate Immune Defense of the Sponge Suberites domuncula against Bacteria Involves a MyD88-dependent Signaling Pathway

Wiens

Korzhev

Krasko

et al. 2005

Journal of Biological Chemistry

168

172

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Sponges (phylum Porifera) are the phylogenetically oldest metazoa; as filter feeders, they are abundantly exposed to marine microorganisms. Here we present data indicating that the demosponge Suberites domuncula is provided with a recognition system for Gramnegative bacteria. The lipopolysaccharide (LPS)-interacting protein was identified as a receptor on the sponge cell surface, which recognizes the bacterial endotoxin LPS. The cDNA was isolated, and the protein (M r 49,937) was expressed. During binding to LPS, the protein dimerizes and interacts with MyD88, which was also identified and cloned. The sponge MyD88 (M r 28,441) is composed of two protein interaction domains, a Toll/interleukin-1 receptor domain (found in MyD88 and in Toll-like receptors) and a death domain (present in MyD88 and interleukin-1 receptor-associated kinase). Northern blot experiments and in situ hybridization studies showed that after LPS treatment, the level of the LPS-interacting protein remains unchanged, whereas MyD88 is strongly up-regulated. A perforin-like molecule (M r 74,171), the macrophage-expressed protein, was identified as an executing molecule of this pathway. This gene is highly expressed after LPS treatment, especially at the surfaces of the animals. The recombinant protein possesses biological activity and eliminates Gram-negative bacteria; it is inactive against Gram-positive bacteria. These data indicate that S. domuncula is provided with an innate immune system against Gramnegative bacteria; the ligand LPS (a pathogen-associated molecular pattern) is recognized by the pattern recognition receptor (LPS-interacting protein), which interacts with MyD88. A signal transduction is established, which results in an elevated expression of MyD88 as well as of the macrophage-expressed protein as an executing protein.

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Section: Discussionmentioning

confidence: 60%

Innate Immune Defense of the Sponge Suberites domuncula against Bacteria Involves a MyD88-dependent Signaling Pathway

Wiens

Korzhev

Krasko

et al. 2005

Journal of Biological Chemistry

168

172

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“…For GRK2, the High Q column simply acts as a prefilter for the removal of contaminants, including other protein kinases (Benovic, Mayor, Staniszewski, Lefkowitz, & Caron, 1987; Gan et al, 2004; Sterne-Marr et al, 2009). This property applies to GRK5 and GRK6, but not to GRK1 or GRK4α, which bind to both cation and anion exchangers (Ohguro et al, 1996; Premont et al, 1996; Sallese et al, 1997).…”

Section: Expression and Purification Of Grksmentioning

confidence: 99%

Expression, Purification, and Analysis of G-Protein-Coupled Receptor Kinases

Sterne‐Marr

Baillargeon

Michalski

et al. 2013

Methods in Enzymology

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G-protein-coupled receptor (GPCR) kinases (GRKs) were first identified based on their ability to specifically phosphorylate activated GPCRs. Although many soluble substrates have since been identified, the chief physiological role of GRKs still remains the uncoupling of GPCRs from heterotrimeric G-proteins by promoting β-arrestin binding through the phosphorylation of the receptor. It is expected that GRKs recognize activated GPCRs through a docking site that not only recognizes the active conformation of the transmembrane domain of the receptor but also stabilizes a more catalytically competent state of the kinase domain. Many of the recent gains in understanding GRK-receptor interactions have been gleaned through biochemical and structural analysis of recombinantly expressed GRKs. Described herein are current techniques and procedures being used to express, purify, and assay GRKs in both in vitro and living cells.

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“…The presence of ionic interactions between GRK2 and its substrate is also supported by the observation that GRK2-mediated receptor phosphorylation is inhibited by salt (30). An additional region that has been implicated in GRK2⅐GPCR interaction is a proline-rich motif just aminoterminal to the nucleotide gate (31). Point mutations in this region not only disrupted the interaction of GRK2 with rhodopsin but also abolished the ability of GRK2 to phosphorylate rhodopsin.…”

Section: Figmentioning

confidence: 99%

Phosphorylation-independent Regulation of Metabotropic Glutamate Receptor 1 Signaling Requires G Protein-coupled Receptor Kinase 2 Binding to the Second Intracellular Loop

Dhami

Babwah

Sterne‐Marr

et al. 2005

Journal of Biological Chemistry

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Metabotropic glutamate receptors (mGluRs) are members of a unique class of G protein-coupled receptors (class III) that include the calcium-sensing and ␥-aminobutyric acid type B receptors. The activity of mGluRs is regulated by second messenger-dependent protein kinases and G protein-coupled receptor kinases (GRKs). The attenuation of both mGluR1a and mGluR1b signaling by GRK2 is phosphorylation-and ␤-arrestin-independent and requires the concomitant association of GRK2 with both the receptor and G␣ q/11 . G protein interactions are mediated, in part, by the mGluR1 intracellular second loop, but the domains required for GRK2 binding are unknown. In the present study, we showed that GRK2 binds to the second intracellular loop of mGluR1a and mGluR1b and also to the mGluR1a carboxyl-terminal tail. Alanine scanning mutagenesis revealed a discrete domain within loop 2 that contributes to GRK2 binding, and the mutation of either lysine 691 or 692 to an alanine within this domain resulted in a loss of GRK2 binding to both mGluR1a and mGluR1b. Mutation of either Lys 691 or Lys 692 prevented GRK2-mediated attenuation of mGluR1b signaling, whereas the mutation of only Lys 692 prevented GRK2-mediated inhibition of mGluR1a signaling. Thus, the mGluR1a carboxyl-terminal tail may also be involved in regulating the signaling of the mGluR1a splice variant. Taken together, our findings indicated that kinase binding to an mGluR1 domain involved in G protein-coupling is essential for the phosphorylation-independent attenuation of signaling by GRK2.Metabotropic glutamate receptors (mGluRs) 1 comprise a family of eight G protein-coupled receptors (GPCRs) that are activated by the excitatory amino acid glutamate and play an important role in regulating neuronal development and synaptic plasticity (1-3). Group I mGluRs (mGluR1 and mGluR5) are coupled via the heterotrimeric G protein G␣ q/11 to the activation of phospholipase C, resulting in the formation of both inositol 1,4,5-triphosphate and diacylglycerol, as well as increases in intracellular Ca 2ϩ concentrations. The alternative gene splicing of mGluR1 generates five carboxyl-terminal domain mGluR1 splice variants (1a, 1b, 1c, 1d, and 1e) (1-4). The G protein-coupling domain for these slice variants is conserved, and it is mediated by amino acid residues localized within the second and third intracellular loops and the membrane proximal region of the mGluR1 carboxyl-terminal tail (4).Similar to most GPCRs, mGluR activity is regulated by serine/threonine protein kinases (5-9). In general, the desensitization of GPCRs involves either phosphorylation by second messenger-dependent protein kinase or phosphorylation by G protein-coupled receptor kinases (GRKs) to promote the binding of arrestin proteins (10, 11). GRK2, GRK4, and GRK5 have each been demonstrated to contribute to the desensitization and internalization of mGluR1 in both heterologous cell cultures and Purkinje cells (5)(6)(7)(12)(13)(14)(15). Although GRK2 and GRK4 promote the ␤-arrestin-dependent internalization of mGlu...

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Involvement of the C-terminal Proline-rich Motif of G Protein-coupled Receptor Kinases in Recognition of Activated Rhodopsin

Cited by 23 publications

References 36 publications

Innate Immune Defense of the Sponge Suberites domuncula against Bacteria Involves a MyD88-dependent Signaling Pathway

Innate Immune Defense of the Sponge Suberites domuncula against Bacteria Involves a MyD88-dependent Signaling Pathway

Expression, Purification, and Analysis of G-Protein-Coupled Receptor Kinases

Phosphorylation-independent Regulation of Metabotropic Glutamate Receptor 1 Signaling Requires G Protein-coupled Receptor Kinase 2 Binding to the Second Intracellular Loop

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