Oligomer formation of histamine H2 receptors expressed in Sf9 and COS7 cells<sup>1</sup>

Fukushima, Yasushi; Asano, Tomohiko; Saitoh, Toshihito; Anai, Motonobu; Funaki, Makoto; Ogihara, Takehide; Katagiri, Hideki; Matsuhashi, Nobuyuki; Yazaki, Yoshio; Sugano, Kentaro

doi:10.1016/s0014-5793(97)00531-0

Cited by 60 publications

(36 citation statements)

References 16 publications

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“…Currently, we do not know the identity of the multiple bands in Sf9 membranes expressing gpH 1 R, but atypical migration of GPCRs in SDS-PAGE has been repeatedly observed (Grü newald et al, 1996;Kelley et al, 2001;Seifert and Wenzel-Seifert, 2001). Because even complex supramolecular structures such as GPCR dimers are preserved in SDS-PAGE (Fukushima et al, 1997;Hebert and Bouvier, 1998;Kelley et al, 2001), it is possible that the different electrophoretic mobilities of hH 1 R and gpH 1 R reflect different GPCR conformations. The different GPCR conformations may be associated with the specific pharmacological properties of H 1 R species isoforms.…”

Section: Discussionmentioning

confidence: 99%

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H₁-Receptor

Seifert¹,

Wenzel-Seifert²,

Bürckstümmer³

et al. 2003

J Pharmacol Exp Ther

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Species isoforms of histamine H 2 -, H 3 -, and H 4 -receptors differ in their pharmacological properties. The study aim was to dissect differences between the human H 1 R (hH 1 R) and guinea pig H 1 R (ghH 1 R). We coexpressed hH 1 R and gpH 1 R with regulators of G-protein signaling in Sf9 insect cells and analyzed the GTPase activity of G q -proteins. Small H 1 R agonists showed similar effects at hH 1 R and gpH 1 R, whereas bulkier 2-phenylhistamines and histaprodifens were up to ϳ10-fold more potent at gpH 1 R than at hH 1 R. Most 2-phenylhistamines and histaprodifens were more efficacious at gpH 1 R than at hH 1 R. Several first-generation H 1 R antagonists were ϳ2-fold, and arpromidine-type H 1 R antagonists up to ϳ10-fold more potent at gpH 1 R than at hH 1 R. [ The Phe-1533 Leu-153/Ile-4333 Val-433 double mutant expressed excellently but only partially changed the pharmacological properties of hH 1 R. Small H 1 R agonists and 2-phenylhistamines interacted differentially with human and guinea pig H 2 R in terms of potency and efficacy, respectively. Our data show the following: 1) there are differences in agonist-and antagonistpharmacology of hH 1 R and gpH 1 R encompassing diverse classes of bulky ligands. These differences may be explained by higher conformational flexibility of gpH 1 R relative to hH 1 R; 2) Phe-153 and Ile-433 are critical for proper folding and expression of hH 1 R; and 3) H 2 R species isoforms distinguish between H 1 R agonists.Histamine serves as a neurotransmitter and autacoid and acts through specific H x Rs designated as H 1 R, H 2 R, H 3 R, and H 4 R, respectively (Hill et al., 1997;Hough, 2001). The H 1 R couples to G q -proteins. Numerous H 1 R agonists and antagonists are known. H 1 R agonists are divided into three classes ( Fig. 1): 1) small agonists (2-4) derived from histamine (1), 2) histamine derivatives with bulkier aromatic substituents at position 2 of the imidazole ring (5-18), and 3) histaprodifens, e.g., compounds 19 to 23 Zingel et al., 1995;Elz et al., 2000). H 1 R agonists are important experimental tools to analyze H 1 R function in cellular and organ systems (Zingel et al., 1995;Hill et al., 1997). H 1 R antagonists are commonly divided into sedating (first-generation, 24-32) and nonsedating (second-generation, 41-45) antagonists (Fig. 2). Today, especially the second-generation H 1 R antagonists are of great importance for the treatment of allergic diseases (Hill et al., 1997). Guanidines 33, 34, and 36 to 39 derived from arpromidine (35) are dual H 2 R agonists/ H 1 R antagonists (Buschauer, 1989).The availability of H x R cDNAs allowed for the comparison of the pharmacological properties of H x R species isoforms in recombinant systems under identical experimental conditions. Such expression studies uncovered species differences This work was supported by the National Institutes of Health COBRE Award 1 P20 RR15563 and matching support from the State of Kansas and the University of Kansas (R.S.), a grant from the Army Research Office (DAAD 19-00-...

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Section: Discussionmentioning

confidence: 99%

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H₁-Receptor

Seifert¹,

Wenzel-Seifert²,

Bürckstümmer³

et al. 2003

J Pharmacol Exp Ther

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“…A number of GPCRs have been shown to be capable of forming homodimeric (Hebert et al, 1996;Fukushima et al, 1997;Xie et al, 1999) or heterodimeric (Jordan and Devi, 1999;Marshall et al, 1999;Xie et al, 1999) structures. It has recently been suggested that the dopamine D3 receptor may also form higher order (dimeric and tetrameric) structures (Nimchinsky et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

The Dopamine D3 Receptor Interacts with Itself and the Truncated D3 Splice Variant D3nf: D3-D3nf Interaction Causes Mislocalization of D3 Receptors

et al. 2000

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We have generated a stable cell line expressing FLAG epitopetagged D3 dopamine receptors and used this cell line to study D3 receptor-protein interactions. To analyze protein interactions, we separately introduced into the stable cell line either D3 receptors carrying an hemagglutinin (HA) epitope tag, or an HA-tagged version of the D3 receptor splice variant D3nf. A combination of confocal laser microscopy and coimmunoprecipitation was used to assay the formation and expression pattern of D3-D3 homodimers or D3-D3nf heterodimers. When coexpressed in HEK 293 cells, FLAG-and HA-tagged D3 receptors exhibited a similar plasma membrane distribution. Using an HA epitope tag-specific antibody, we coimmunoprecipitated HA-and FLAG-tagged D3 receptors, suggesting that D3 receptors are capable of forming homodimers. Epitope-tagged D3nf polypeptides exhibited a markedly different cellular distribution than D3 receptors. When expressed in HEK 293 cells, either alone or in combination with FLAG-tagged D3 receptors, D3nf exhibited a punctate perinuclear distribution. When D3nf was introduced into the stable D3-expressing cell line, D3 receptors were no longer visualized at the plasma membrane. Instead, D3 and D3nf showed a similar, predominantly cytosolic, localization. Using the HA-specific antibody, we were able to coimmunoprecipitate D3 and D3nf polypeptides from transfected cells. These data suggest the existence of physical interaction between D3 and D3nf. This interaction appears to result in the mislocalization of D3 receptors from the plasma membrane to an intracellular compartment, a finding that could be of significance in the etiology of schizophrenia.Dopamine neurotransmission in mammalian brain is mediated by a cohort of receptors that are members of the superfamily of G protein-coupled receptors (GPCRs). In humans, five dopamine receptor subtypes (D1-D5) have been identified by molecular cloning (reviewed in Missale et al., 1998). The five dopamine receptors have been grouped into two subfamilies based upon sequence homologies and pharmacologic profiles. The D1 class of dopamine receptors is comprised of the D1 and D5 receptor subtypes. These receptors are expressed at high levels in cerebral cortex and are coupled to stimulatory subsets of heterotrimeric G proteins. The D2 class of dopamine receptors, consisting of the D2, D3, and D4 subtypes, is coupled to inhibitory subsets of G proteins and is a major target of antipsychotic drugs.Among the D2 class of dopamine receptors, the D3 receptor is distinctive in that it is distributed preferentially in limbic areas of the brain (nucleus accumbens, olfactory tubercle, islands of Calleja, and hippocampus) thought to control cognitive and emotional aspects of behavior (Bouthenet et al., 1991). The D3 receptor has also been shown to bind most antipsychotic drugs, both typical and atypical, with high affinity (Levant, 1997). Because of its anatomic distribution and interaction with antipsychotic drugs, the D3 dopamine receptor has been suggested to play a role i...

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“…Recent studies have shown that G-protein-coupled receptors (GPCRs) 1 may form dimers or higher order oligomers (1)(2)(3)(4)(5)(6)(7). This has led to some re-evaluation of the mechanisms thought to be involved in GPCR function.…”

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confidence: 99%

Oligomerization of G-protein-coupled Receptors Shown by Selective Co-immunoprecipitation

Salim

Fenton

Bacha

et al. 2002

Journal of Biological Chemistry

108

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Recent studies have shown that G-protein-coupled receptors (GPCRs) can assemble as high molecular weight homo-and hetero-oligomeric complexes. This can result in altered receptor-ligand binding, signaling, or intracellular trafficking. We have co-transfected HEK-293 cells with differentially epitope-tagged GPCRs from different subfamilies and determined whether oligomeric complexes were formed by co-immunoprecipitation and immunoblot analysis. This gave the surprising result that the 5HT 1A receptor was capable of forming heterooligomers with all GPCRs tested including the 5HT 1B , 5HT 1D , EDG 1 , EDG 3 , GPR 26 , and GABA B2 receptors. The testing of other GPCR combinations showed similar results with hetero-oligomer formation occurring for the 5HT 1D with the 5HT 1B and EDG 1 receptor. Control studies showed that these complexes were present in cotransfected cells before the time of lysis and that the hetero-oligomers were comprised of GPCRs at discrete stoichiometries. These findings suggest that GPCRs have a natural tendency to form oligomers when cotransfected into cells. Future studies should therefore investigate the presence and physiological role of GPCR hetero-oligomers in cells in which they are endogenously expressed.

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Oligomer formation of histamine H2 receptors expressed in Sf9 and COS7 cells¹

Cited by 60 publications

References 16 publications

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H₁-Receptor

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H₁-Receptor

The Dopamine D3 Receptor Interacts with Itself and the Truncated D3 Splice Variant D3nf: D3-D3nf Interaction Causes Mislocalization of D3 Receptors

Oligomerization of G-protein-coupled Receptors Shown by Selective Co-immunoprecipitation

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Oligomer formation of histamine H2 receptors expressed in Sf9 and COS7 cells1

Cited by 60 publications

References 16 publications

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H1-Receptor

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H1-Receptor

The Dopamine D3 Receptor Interacts with Itself and the Truncated D3 Splice Variant D3nf: D3-D3nf Interaction Causes Mislocalization of D3 Receptors

Oligomerization of G-protein-coupled Receptors Shown by Selective Co-immunoprecipitation

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Oligomer formation of histamine H2 receptors expressed in Sf9 and COS7 cells¹

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H₁-Receptor

Multiple Differences in Agonist and Antagonist Pharmacology between Human and Guinea Pig Histamine H₁-Receptor