Structure Of The Adrenergic And Related Receptors

O'dowd, B.

doi:10.1146/annurev.neuro.12.1.67

Cited by 18 publications

(18 citation statements)

References 21 publications

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“…The final 220 residues of the protein, -45% of the sequence, are primarily hydrophilic. This topology is characteristic of the superfamily of G-protein-coupled receptors, such as the adrenergic receptors (O'Dowd et al 1989) and the rhodopsins (Findlay and Pappin 1986;Hargrave 1986). Similar to the proposed structure of these receptors, cAR3 would have an extracellular amino terminus and a long cytoplasmic carboxyl terminus.…”

Section: Car3 Sequencementioning

confidence: 99%

“…The cytoplasmic carboxy-terminal domain of cAR3 contains multiple serine and threonine residues, which are the sites of ligand-induced phosphorylation in the B-adrenergic receptor {O'Dowd et al 1989), rhodopsin {Kuhn and Dreyer 19721, and cAR1 (Vaughan and Devreotes 1988; D. Hereld, R. Vaughan, and P. Devreotes, in prep.I. An additional 4 serine and 3 threonine residues are present in the intracellular loop between the fifth and sixth putative transmembrane regions.…”

Section: Sequence Comparisonmentioning

confidence: 99%

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Identification and targeted gene disruption of cAR3, a cAMP receptor subtype expressed during multicellular stages of Dictyostelium development.

Johnson¹,

Saxe²,

Gollop³

et al. 1993

Genes Dev.

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Extracellular cAMP acts through cell-surface receptors to coordinate the developmental program of Dictyostelium.A cAMP receptor (cAR1), which is expressed during early aggregation, has been cloned and sequenced previously. We have identified a new receptor subtype, cAR3, that has -56% and 69% amino acid identity with cAR1 and cAR2, respectively, cAR1, cAR2, or cAR3 expressed from plasmid in growing Dictyostelinm cells can be photoaffinity labeled with 8-N3132p]cAMP and phosphorylated when stimulated with cAMP. cAR3 RNA was not present during growth but appeared during late aggregation. Its expression peaked at 9 hr and then fell to a reduced level that was maintained until culmination. The expression of cAR3 protein followed a similar pattern, but with a 3-hr lag, and reached a maximum at the mound stage. In contrast, cAR1 protein was expressed predominantly during early aggregation and at low levels during later stages. At their respective peaks of expression, there were -5 x 103 cAR3 sites per cell compared with -7 x 104 cAR1 sites per cell. The cAR3 gene was disrupted by homologous recombination in several different parental cell lines. Surprisingly, the car3-cell lines display no obvious phenotype.

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Section: Car3 Sequencementioning

confidence: 99%

Section: Sequence Comparisonmentioning

confidence: 99%

Identification and targeted gene disruption of cAR3, a cAMP receptor subtype expressed during multicellular stages of Dictyostelium development.

Johnson¹,

Saxe²,

Gollop³

et al. 1993

Genes Dev.

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“…In all cases, G-protein-linked receptors have been found to be composed of single unit. Significant sequence homologies are found among all G-protein linked receptors, and they are translated into a con served seven-transmembrane-spanning three-dimension al structure (12,13). Another common structural fea ture of these receptors is related to the presence of potential N-linked glycosylation sites at the amino ter minal (13).…”

Section: Discussionmentioning

confidence: 99%

Isolation of Substance P Binding Protein from Rat Brain.

et al. 1992

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ABSTRACT-Substance P (SP) binding protein of rat brain was solubilized by digitonin. The solubil ized proteins were then purified by sequential gel filtration, concanavalin A lectin Sepharose, and SP affinity chromatography. The calculated molecular weight of this purified SP binding protein was 76-74 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The rabbits were immunized with the purified protein and resulting polyclonal anti-sera were tested. The immune serum significantly in hibited [3H]SP binding to the 3- [(3-cholamidopropyl)dimethyl ammonio]-1-propane sulfonate solubilized membrane fractions from rat brain, whereas pre-bleed antiserum failed to inhibit the binding. This polyclonal antibody also inhibited the activity of 45 Ca influx into astroglioma cells stimulated by SP, but does not inhibit that stimulated by histamine. Furthermore, this polyclonal antibody recognized the 76 74 kDa band as assessed by Western blotting. These data strongly suggest that this polyclonal antibody could recognize a part of the natural SP receptor site.Keywords: Substance P, Binding protein, Brain (rat), Polyclonal antibody (substance P), Affinity chromatography (substance P) Purification and characterization of the substance P (SP) receptor are important in understanding the molecular basis for the biological action between SP and its receptor. Unfortunately, there are no reports about the isolation of the SP receptor protein. Howev er, the mRNA for the SP receptor was recently cloned and the amino acid sequence was determined (1). The molecular cloning and characterization of rat SP recep tor have been reported by Hershey and Krause (2). The sequence showed the presence of N-glycosylation sites and hydrophobic membrane-spanning segments. The SP receptor belongs to the family of G-protein coupled receptors and affects their functions by mod ulating inositol phosphate/calcium second messengers (3). The isolation of several other G-protein-coupled receptors, for example a and /8-adrenergic receptors and muscarine receptors, has been achieved by affinity chromatography. Therefore, the present study was de signed to isolate SP binding protein with carbohydrate moieties by affinity chromatography and to make a polyclonal antibody for this protein. Using the poly clonal antibody, we could demonstrate a decrease in the SP-induced biological activity by covering the SP re ceptor with the polyclonal antibody. MATERIALS AND METHODSMembrane preparation and solubilization Male Wistar rats (180-250g) were used. Cerebral cortical membranes were prepared by the method of Nakata et al. (4). Immediately, after decapitation, the cerebral cortices were homogenized in ice-cold 0.32 M sucrose with a glass-Teflon homogenizer. Homogenates were centrifuged at 1,000 X g for 10 min to obtain the supernatants. These supernatants were then centrifuged at 17,000 X g for 20 min, and the pellets obtained were washed three times and used immediately for the solu bilization experiments. For solubilization, the pellets were suspended in ...

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“…Structural characteristics that are shared by most of the G protein-linked receptors The last extracellular loop is generally small. This is thought to facilitate the close apposition of the cytoplasmic loop between TM5 and TM6 and the cytoplasmic domain carboxy-terminal to TM7 (O'Dowd et al 1989). Evidence suggests that these cytoplasmic regions are directly involved in G protein interactions (Strader et al 1987;Dixon et al 1988;O'Dowd et al 1988).…”

Section: Genes and Developmentmentioning

confidence: 99%

“…Evidence suggests that these cytoplasmic regions are directly involved in G protein interactions (Strader et al 1987;Dixon et al 1988;O'Dowd et al 1988). (2) The third cytoplasmic loop connecting TM5 and TM6 contains positively charged amino acids that can be arranged along one side of an amphipathic ahelix; this structure is thought to activate G proteins (O'Dowd et al 1989). (3) A cysteine is found in each of the first two short extracellular loops.…”

Section: Genes and Developmentmentioning

confidence: 99%

Induction of cell fate in the Drosophila retina: the bride of sevenless protein is predicted to contain a large extracellular domain and seven transmembrane segments.

et al. 1990

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Previous genetic mosaic studies established that expression of the Drosophila bride of sevenless (boss) gene is required in photoreceptor neuron R8 for the development of photoreceptor neuron R7. This led to the proposal that boss encodes or regulates an R7-specific inductive cue. We have identified the boss gene based on small deletions in mutant alleles and sequenced both cDNAs and corresponding genomic regions. One P element and three X-ray-induced boss alleles show different deletions in the gene ranging in size from 2 to 23 bp, each causing frameshifts leading to premature termination of translation. The boss gene encodes a protein of 896 amino acids with a putative amino-terminal signal sequence, a large extracellular region of 498 amino acids, and seven potential transmembrane domains followed by a carboxy-terminal cytoplasmic tail of 115 amino acids. The putative membrane localization of the boss protein is consistent with a model in which direct interaction between the boss and sevenless proteins specifies R7 cell fate. Another model in which the boss protein functions as a receptor is proposed based on its similarity to the G protein-linked family of membrane receptors.

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Structure Of The Adrenergic And Related Receptors

Cited by 18 publications

References 21 publications

Identification and targeted gene disruption of cAR3, a cAMP receptor subtype expressed during multicellular stages of Dictyostelium development.

Identification and targeted gene disruption of cAR3, a cAMP receptor subtype expressed during multicellular stages of Dictyostelium development.

Isolation of Substance P Binding Protein from Rat Brain.

Induction of cell fate in the Drosophila retina: the bride of sevenless protein is predicted to contain a large extracellular domain and seven transmembrane segments.

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