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1.
The inhibitor protein of the cyclic AMP-dependent protein kinase-catalytic subunit interaction. Composition of multiple complexes. 总被引:1,自引:0,他引:1
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It has been previously demonstrated that the combination of pure preparations of the inhibitor protein of the cyclic AMP-dependent protein kinase and the catalytic subunit of this enzyme resulted in the formation of multiple complexes [Van Patten, Fletcher & Walsh (1986) J. Biol. Chem. 261, 5514-5523]. In the present study it is demonstrated that these multiple species occur because the bovine heart protein kinase preparation contains multiple forms of catalytic subunit [Kinzel, Hotz, König, Gagelmann, Pyerin, Reed, Köbler, Hofmann, Obst, Gensheimer, Goldblatt & Shaltiel (1987) Arch. Biochem. Biophys. 253, 341-349]. 相似文献
2.
Lee SW Bonnah RA Higashi DL Atkinson JP Milgram SL So M 《The Journal of cell biology》2002,156(6):951-957
The Neisseria type IV pilus promotes bacterial adhesion to host cells. The pilus binds CD46, a complement-regulatory glycoprotein present on nucleated human cells (Källström et al., 1997). CD46 mutants with truncated cytoplasmic tails fail to support bacterial adhesion (Källström et al., 2001), suggesting that this region of the molecule also plays an important role in infection. Here, we report that infection of human epithelial cells by piliated Neisseria gonorrhoeae (GC) leads to rapid tyrosine phosphorylation of CD46. Studies with wild-type and mutant tail fusion constructs demonstrate that Src kinase phosphorylates tyrosine 354 in the Cyt2 isoform of the CD46 cytoplasmic tail. Consistent with these findings, infection studies show that PP2, a specific Src family kinase inhibitor, but not PP3, an inactive variant of this drug, reduces the ability of epithelial cells to support bacterial adhesion. Several lines of evidence point to the role of c-Yes, a member of the Src family of nonreceptor tyrosine kinases, in CD46 phosphorylation. GC infection causes c-Yes to aggregate in the host cell cortex beneath adherent bacteria, increases binding of c-Yes to CD46, and stimulates c-Yes kinase activity. Finally, c-Yes immunoprecipitated from epithelial cells is able to phosphorylate the wild-type Cyt2 tail but not the mutant derivative in which tyrosine 354 has been substituted with alanine. We conclude that GC infection leads to rapid tyrosine phosphorylation of the CD46 Cyt2 tail and that the Src kinase c-Yes is involved in this reaction. Together, the findings reported here and elsewhere strongly suggest that pilus binding to CD46 is not a simple static process. Rather, they support a model in which pilus interaction with CD46 promotes signaling cascades important for Neisseria infectivity. 相似文献
3.
A second gene for autosomal dominant Möbius syndrome is localized to chromosome 10q, in a Dutch family.
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H T Verzijl B van den Helm B Veldman B C Hamel L P Kuyt G W Padberg H Kremer 《American journal of human genetics》1999,65(3):752-756
Möbius syndrome (MIM 157900) consists of a congenital paresis or paralysis of the VIIth (facial) cranial nerve, frequently accompanied by dysfunction of other cranial nerves. The abducens nerve is typically affected, and often, also, the hypoglossal nerve. In addition, orofacial and limb malformations, defects of the musculoskeletal system, and mental retardation are seen in patients with Möbius syndrome. Most cases are sporadic, but familial recurrence can occur. Different modes of inheritance are suggested by different pedigrees. Genetic heterogeneity of Möbius syndrome has been suggested by cytogenetic studies and linkage analysis. Previously, we identified a locus on chromosome 3q21-22, in a large Dutch family with Möbius syndrome consisting essentially of autosomal dominant asymmetric bilateral facial paresis. Here we report linkage analysis in a second large Dutch family with autosomal dominant inherited facial paresis. After exclusion of >90% of the genome, we identified the locus on the long arm of chromosome 10 in this family, demonstrating genetic heterogeneity of this condition. The reduced penetrance suggests that at least some of the sporadic cases might be familial. 相似文献
4.
EM Putz M Prchal-Murphy OA Simma F Forster X Koenig H Stockinger RP Piekorz M Freissmuth M Müller V Sexl E Zebedin-Brandl 《PloS one》2012,7(7):e40852
Background
PI3Kδ is a lipid kinase of the phosphoinositide 3-kinase class 1A family and involved in early signaling events of leukocytes regulating proliferation, differentiation and survival. Currently, several inhibitors of PI3Kδ are under investigation for the treatment of hematopoietic malignancies. In contrast to the beneficial effect of inhibiting PI3Kδ in tumor cells, several studies reported the requirement of PI3Kδ for the function of immune cells, such as natural killer and T helper cells. Cytotoxic T lymphocytes (CTLs) are essential for tumor surveillance. The scope of this study is to clarify the potential impact of PI3Kδ inhibition on the function of CTLs with emphasis on tumor surveillance.Principal Findings
PI3Kδ-deficient mice develop significantly bigger tumors when challenged with MC38 colon adenocarcinoma cells. This defect is accounted for by the fact that PI3Kδ controls the secretory perforin-granzyme pathway as well as the death-receptor pathway of CTL-mediated cytotoxicity, leading to severely diminished cytotoxicity against target cells in vitro and in vivo in the absence of PI3Kδ expression. PI3Kδ-deficient CTLs express low mRNA levels of important components of the cytotoxic machinery, e.g. prf1, grzmA, grzmB, fasl and trail. Accordingly, PI3Kδ-deficient tumor-infiltrating CTLs display a phenotype reminiscent of naïve T cells (CD69lowCD62Lhigh). In addition, electrophysiological capacitance measurements confirmed a fundamental degranulation defect of PI3Kδ−/− CTLs.Conclusion
Our results demonstrate that CTL-mediated tumor surveillance is severely impaired in the absence of PI3Kδ and predict that impaired immunosurveillance may limit the effectiveness of PI3Kδ inhibitors in long-term treatment. 相似文献5.
Nicolas X. Tritsch Ying-Xin Zhang Graham Ellis-Davies Dwight E. Bergles 《Purinergic signalling》2010,6(2):155-166
The developing cochlea of mammals contains a large group of columnar-shaped cells, which together form a structure known as Kölliker’s organ. Prior to the onset of hearing, these inner supporting cells periodically release adenosine 5′-triphosphate (ATP), which activates purinergic receptors in surrounding supporting cells, inner hair cells and the dendrites of primary auditory neurons. Recent studies indicate that purinergic signaling between inner supporting cells and inner hair cells initiates bursts of action potentials in auditory nerve fibers before the onset of hearing. ATP also induces prominent effects in inner supporting cells, including an increase in membrane conductance, a rise in intracellular Ca2+, and dramatic changes in cell shape, although the importance of ATP signaling in non-sensory cells of the developing cochlea remains unknown. Here, we review current knowledge pertaining to purinergic signaling in supporting cells of Kölliker’s organ and focus on the mechanisms by which ATP induces changes in their morphology. We show that these changes in cell shape are preceded by increases in cytoplasmic Ca2+, and provide new evidence indicating that elevation of intracellular Ca2+ and IP3 are sufficient to initiate shape changes. In addition, we discuss the possibility that these ATP-mediated morphological changes reflect crenation following the activation of Ca2+-activated Cl− channels, and speculate about the possible functions of these changes in cell morphology for maturation of the cochlea. 相似文献
6.
Cecilia Y. Cheng Jie Yang Susan S. Taylor Donald K. Blumenthal 《The Journal of biological chemistry》2009,284(51):35916-35925
The catalytic (C) and regulatory (R) subunits of protein kinase A are exceptionally dynamic proteins. Interactions between the R- and C-subunits are regulated by cAMP binding to the two cyclic nucleotide-binding domains in the R-subunit. Mammalian cells express four different isoforms of the R-subunit (RIα, RIβ, RIIα, and RIIβ) that all interact with the C-subunit in different ways. Here, we investigate the dynamic behavior of protein complexes between RIα and C-subunits using small angle x-ray scattering. We show that a single point mutation in RIα, R333K (which alters the cAMP-binding properties of Domain B) results in a compact shape compared with the extended shape of the wild-type R·C complex. A double mutant complex that disrupts the interaction site between the C-subunit and Domain B in RIα, RIαABR333K·C(K285P), results in a broader P(r) curve that more closely resembles the P(r) profiles of wild-type complexes. These results together suggest that interactions between RIα Domain B and the C-subunit in the RIα·C complex involve large scale dynamics that can be disrupted by single point mutations in both proteins. In contrast to RIα·C complexes. Domain B in the RIIβ·C heterodimer is not dynamic and is critical for both inhibition and complex formation. Our study highlights the functional differences of domain dynamics between protein kinase A isoforms, providing a framework for elucidating the global organization of each holoenzyme and the cross-talk between the R- and C-subunits. 相似文献
7.
G. Gregory Neely Shuan Rao Michael Costigan Norbert Mair Ildiko Racz Giedre Milinkeviciute Arabella Meixner Swetha Nayanala Robert S. Griffin Inna Belfer Feng Dai Shad Smith Luda Diatchenko Stefano Marengo Bernhard J. Haubner Maria Novatchkova Dustin Gibson William Maixner J. Andrew Pospisilik Emilio Hirsch Ian Q. Whishaw Andreas Zimmer Vaijayanti Gupta Junko Sasaki Yasunori Kanaho Takehiko Sasaki Michaela Kress Clifford J. Woolf Josef M. Penninger 《PLoS genetics》2012,8(12)
The ability to perceive noxious stimuli is critical for an animal''s survival in the face of environmental danger, and thus pain perception is likely to be under stringent evolutionary pressure. Using a neuronal-specific RNAi knock-down strategy in adult Drosophila, we recently completed a genome-wide functional annotation of heat nociception that allowed us to identify α2δ3 as a novel pain gene. Here we report construction of an evolutionary-conserved, system-level, global molecular pain network map. Our systems map is markedly enriched for multiple genes associated with human pain and predicts a plethora of novel candidate pain pathways. One central node of this pain network is phospholipid signaling, which has been implicated before in pain processing. To further investigate the role of phospholipid signaling in mammalian heat pain perception, we analysed the phenotype of PIP5Kα and PI3Kγ mutant mice. Intriguingly, both of these mice exhibit pronounced hypersensitivity to noxious heat and capsaicin-induced pain, which directly mapped through PI3Kγ kinase-dead knock-in mice to PI3Kγ lipid kinase activity. Using single primary sensory neuron recording, PI3Kγ function was mechanistically linked to a negative regulation of TRPV1 channel transduction. Our data provide a systems map for heat nociception and reinforces the extraordinary conservation of molecular mechanisms of nociception across different species. 相似文献
8.
Bacteriolytic enzymes from Staphylococcus aureus. Properties of the endo-β-N-acetylglucosaminidase 总被引:4,自引:0,他引:4
T. Wadstr?m 《The Biochemical journal》1970,120(4):745-752
An extracellular bacteriolytic endo-β-N-acetylglucosaminidase has been purified and its specificity of action has been investigated (Wadström & Hisatsune, 1970a,b). Some enzymic properties, such as optimum pH for enzyme activity on whole cells and cell walls of Micrococcus lysodeikticus and Staphylococcus aureus and optimum pH for stability, have been studied. The activity was maximum in 0.05m-tris–hydrochloric acid buffer, pH7.0. A higher ionic strength inhibited cell-wall hydrolysis. Since the crude and purified enzymes were found to be unstable on storage, the stabilizing and inhibiting effects of several compounds were investigated. Several heavy metal ions inactivated the enzyme at very low concentrations. Thiol compounds stabilized and thiol-reacting compounds partly inhibited the activity. Crude and purified glucosaminidase was found to be heat-stable at acidic pH and unstable at alkaline pH as previously found for several lysozymes (endo-β-N-acetylmuramidases). Other properties of the staphylococcal enzyme and hen''s-egg-white lysozyme have been compared, since the modes of action of the two are quite similar (Wadström & Hisatsune, 1970b). 相似文献
9.
Thioamides, urea and thiourea derivatives of 2,6-pyridinedicarbonyl dichloride, isophthaloyl dichloride and terephthaloyl dichloride have been synthesized. These ligands have been incorporated in dinuclear rhenium(I) diimine tricarbonyl complexes and the anion recognition properties of these complexes have been studied by luminescence, UV-Vis and 1H NMR spectroscopic methods. The complexes act as receptors for anions via hydrogen bonding and electrostatic interactions. The anion sensing properties of the complexes are compared to earlier amide-based dinuclear rhenium(I) tricarbonyl complexes. 相似文献
10.
Leonardus M. I. Koharudin Sireesha Kollipara Christopher Aiken Angela M. Gronenborn 《The Journal of biological chemistry》2012,287(40):33796-33811
Oscillatoria agardhii agglutinin homolog (OAAH) proteins belong to a recently discovered lectin family. All members contain a sequence repeat of ∼66 amino acids, with the number of repeats varying among different family members. Apart from data for the founding member OAA, neither three-dimensional structures, information about carbohydrate binding specificities, nor antiviral activity data have been available up to now for any other members of the OAAH family. To elucidate the structural basis for the antiviral mechanism of OAAHs, we determined the crystal structures of Pseudomonas fluorescens and Myxococcus xanthus lectins. Both proteins exhibit the same fold, resembling the founding family member, OAA, with minor differences in loop conformations. Carbohydrate binding studies by NMR and x-ray structures of glycan-lectin complexes reveal that the number of sugar binding sites corresponds to the number of sequence repeats in each protein. As for OAA, tight and specific binding to α3,α6-mannopentaose was observed. All the OAAH proteins described here exhibit potent anti-HIV activity at comparable levels. Altogether, our results provide structural details of the protein-carbohydrate interaction for this novel lectin family and insights into the molecular basis of their HIV inactivation properties. 相似文献
11.
Mohamed Hussain Syed Abuthakir Munirah Abdullah Al-Dosary Ashraf Atef Hatamleh Hissah Abdulrahman Alodaini P. Perumal Muthusamy Jeyam 《Saudi Journal of Biological Sciences》2022,29(5):3899
Trichophyton rubrum is one of the major disease causing pathogens in human; mainly it causes tinea pedis, tinea cruris and tinea corporis. Cytochrome P450 which considered to be an important protein that can impact ergosterol biosynthesis pathway. B. aegyptiaca is rich source of secondary metabolites with tremendous medicinal values and it has sweet pulp, leaves with spine, strong seed and oily kernel. The epicarp of the fruit was taken for this study to inhibit T. rubrum using in vitro and in silico techniques. The epicarp portion was extracted using various solvents and water. The anti-dermatophytic activity on T. rubrum of these extracts was assessed utilizing poison plate technique with 5 individual concentrations. The fractioned chloroform extract of epicarp had fully inhibited the growth of T. rubrum at 3 mg/ml. Further, the chloroform extract was subjected to LC-MS analysis, in total, 40 compounds were elucidated. Then, the derived compounds were included for predicting ADMETox properties using Qikprop module. From the analysis 40 compounds were identified to be eligible for docking process. Then the desirable compounds, drug Ketoconazole were subjected to docking analysis using Glide module of Schrödinger. It shows that Platyphylloside has better docking result than other compounds and drug Ketoconazole. Further, MD simulation was carried out for Ketoconazole-Cyp450 and Platyphylloside-CYP450 complexes using Desmond, Schrödinger. MD simulation study also confirmed that the Platyphylloside-CYP450 complex more stable. This study suggests that Platyphylloside may act as potential inhibitor and it could be further subjected to experimental analysis to inhibit the T. rubrum growth. 相似文献
12.
Dian Spakman Julia A M Bakx Andreas
S Biebricher Erwin J G Peterman Gijs J L Wuite Graeme
A King 《Nucleic acids research》2021,49(10):5470
Topoisomerases are essential enzymes that regulate DNA topology. Type 1A family topoisomerases are found in nearly all living organisms and are unique in that they require single-stranded (ss)DNA for activity. These enzymes are vital for maintaining supercoiling homeostasis and resolving DNA entanglements generated during DNA replication and repair. While the catalytic cycle of Type 1A topoisomerases has been long-known to involve an enzyme-bridged ssDNA gate that allows strand passage, a deeper mechanistic understanding of these enzymes has only recently begun to emerge. This knowledge has been greatly enhanced through the combination of biochemical studies and increasingly sophisticated single-molecule assays based on magnetic tweezers, optical tweezers, atomic force microscopy and Förster resonance energy transfer. In this review, we discuss how single-molecule assays have advanced our understanding of the gate opening dynamics and strand-passage mechanisms of Type 1A topoisomerases, as well as the interplay of Type 1A topoisomerases with partner proteins, such as RecQ-family helicases. We also highlight how these assays have shed new light on the likely functional roles of Type 1A topoisomerases in vivo and discuss recent developments in single-molecule technologies that could be applied to further enhance our understanding of these essential enzymes. 相似文献
13.
The excellence of Károly Sajó as a researcher into Hungary’s natural history has been undeservedly neglected. Yet he did lasting work, especially in entomology, and a number of his discoveries and initiatives were before their time.Born in 1851 in Győr, he received his secondary education there and went to Pest University. He taught in a grammar school in 1877–88 before spending seven years as an entomologist at the National Phylloxera Experimental Station, later the Royal Hungarian State Entomological Station. Pensioned off at his own request in 1895, he moved to Őrszentmiklós, where he continued making entomological observations on his own farm and wrote the bulk of his published materials: almost 500 longer or shorter notes, articles and books, mainly on entomological subjects.Sajó was among the first in the world to publish in 1896 a study of how the weather affects living organisms, entitled Living Barometers. His Sleep in Insects, which appeared in the same year, described his discovery, from 1895 observations of the red turnip beetle, Entomoscelis adonidis (Pallas, 1771), of aestivation in insects – in present-day terms diapause.It was a great loss to universal entomology when Sajó ceased publishing about 25 years before his death. His unpublished notes, with his library and correspondence, were destroyed in the World War II. His surviving insect collection is now kept in the Hungarian Natural History Museum, Budapest. 相似文献
14.
Aliny Pereira Lima Flávia Castro Pereira Marcio Aurelio Pinheiro Almeida Francyelli Mariana Santos Mello Wanessa Carvalho Pires Thallita Monteiro Pinto Flávia Karina Delella Sérgio Luis Felisbino Virtudes Moreno Alzir Azevedo Batista Elisangela de Paula Silveira-Lacerda 《PloS one》2014,9(10)
Over the past several decades, much attention has been focused on ruthenium complexes in antitumor therapy. Ruthenium is a transition metal that possesses several advantages for rational antitumor drug design and biological applications. In the present study, five ruthenium complexes containing amino acids were studied in vitro to determine their biological activity against sarcoma-180 tumor cells. The cytotoxicity of the complexes was evaluated by an MTT assay, and their mechanism of action was investigated. The results demonstrated that the five complexes inhibited the growth of the S180 tumor cell line, with IC50 values ranging from 22.53 µM to 50.18 µM, and showed low cytotoxicity against normal L929 fibroblast cells. Flow cytometric analysis revealed that the [Ru(gly)(bipy)(dppb)]PF6 complex (2) inhibited the growth of the tumor cells by inducing apoptosis, as evidenced by an increased number of Annexin V-positive cells and G0/G1 phase cell cycle arrest. Further investigation showed that complex 2 caused a loss of mitochondrial membrane potential; activated caspases 3, caspase-8, and caspase-9 and caused a change in the mRNA expression levels of caspase 3, caspase-9 as well as the bax genes. The levels of the pro-apoptotic Bcl-2 family protein Bak were increased. Thus, we demonstrated that ruthenium amino acid complexes are promising drugs against S180 tumor cells, and we recommend further investigations of their role as chemotherapeutic agents for sarcomas. 相似文献
15.
16.
Egg Membrane Lytic Activity of Sperm Extract and its Significance in Relation to Sperm Entry in Hydroides hexagonus (Annelida) 总被引:4,自引:4,他引:0
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Previous electron microscope studies indicated that the individual spermatozoön of Hydroides hexagonus forms a hole in the vitelline membrane by means of lysis. Other observations established that the hole is real, being visible in living material during sperm entry. During the present investigation sea water extracts from frozen-thawed sperm were tested for lytic effect on the membrane. In normal living eggs the membrane appears as a single thick envelope, but in electron micrographs of sections it is seen to consist of a narrow outer border layer, a wide principal or middle layer, and a narrow inner border layer. After immersion in sperm extract the outer border layer elevates but does not dissolve, the middle layer liquefies and disappears, and the inner border layer seems not to change. This is interpreted as lysis of the middle layer. The extract exerted the same effect on fertilized and unfertilized eggs. In electron micrographs the sections treated with extract greatly resemble that part of the membrane which has been penetrated by the individual spermatozoön. It is concluded that the individual spermatozoön, too, exerts a lytic effect. Together, the present and two earlier studies are considered clearly to demonstrate that in Hydroides the individual spermatozoön does indeed make an entry hole in the egg membrane by applying lytic material to that part of the membrane in its own vicinity. 相似文献
17.
Stimulation of polyoma DNA replication in isolated nucleoprotein complexes by factors from Drosophila embryos 总被引:2,自引:0,他引:2
Nucleoprotein complexes containing both form 1 and replicative intermediates of polyoma DNA prepared from nuclei of virus-infected mouse fibroblasts retain a limited ability to elongate progeny strands of the replicative intermediates. Compared to isolated nuclei, both the rate and the extent of strand elongation is greatly decreased. The isolated complexes synthesize initiator RNA and start new Okazaki fragments, but are deficient in the joining of these fragments. Addition of small amounts of an extract from 16 hours old Drosophila embryos corrects the deficiencies. The stimulatory activity of the extract can be partially purified and has been separated into two fractions by chromatography on Sepharose 6B. With immunological techniques we demonstrate that the mouse DNA polymerase-α, tightly bound to the complexes, is responsible for DNA strand elongation.The Drosophila α-polymerase present in one of the two fractions purified on Sepharose 6B cannot substitute for the mouse enzyme. The stimulatory activity of the Drosophila fractions is thus not due to α-polymerase. 相似文献
18.
Formation of Sperm Entry Holes in the Vitelline Membrane of Hydroides hexagonus (Annelida) and Evidence of their Lytic Origin 总被引:3,自引:3,他引:0
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Electron micrographs of inseminated eggs of Hydroides hexagonus previously had shown that in the immediate vicinity of the penetrating spermatozoön a small portion of the vitelline membrane regularly was absent, and it had been suggested that this area was a hole made by lytic activity of the individual spermatozoön during the course of its passage through the membrane. This deduction would receive support if it could be established that a sperm entry hole does form in living material. During the present study a hole repeatedly observed and photographed in the membrane of living eggs was found to arise as the spermatozoön penetrated the membrane. Gently compressed eggs formed exovates only through this hole. The holes, and exovates, were not found except at sperm entry sites. It was concluded that this hole is the counterpart of the area from which the membrane is absent in the electron micrographs cited above, and that the spermatozoön makes this hole. In an electron micrograph two spermatozoa which had penetrated the membrane at separate but closely neighboring points now occupy a single hole. It is argued that if each spermatozoön had displaced the membrane mechanically to make its hole, then there should be two holes, with a partition of membrane between them, but if each had eroded the membrane by applying lysin, a single hole should have formed as the eroded areas expanded and finally merged into one. The latter view agrees with the facts of the electron micrograph. It is concluded that lysis is the most probable means by which the individual spermatozoön makes its hole. 相似文献
19.
Ikuo Masuho Jeremy Celver Abraham Kovoor Kirill A. Martemyanov 《The Journal of biological chemistry》2010,285(7):4781-4787
The R7 subfamily of RGS proteins critically regulates neuronal G protein-signaling pathways that are essential for vision, nociception, motor coordination, and reward processing. A member of the R7 RGS family, RGS11, is a GTPase-accelerating protein specifically expressed in retinal ON-bipolar cells where it forms complexes with the atypical G protein β subunit, Gβ5, and transmembrane protein R9AP. Association with R9AP has been shown to be critical for the proteolytic stability of the complex in the retina. In this study we report that R9AP can in addition stimulate the GTPase-accelerating protein activity of the RGS11·Gβ5 complex at Gαo. Single turnover GTPase assays reveal that R9AP co-localizes RGS11·Gβ5 and Gαo on the membrane and allosterically potentiates the GTPase-accelerating function of RGS11·Gβ5. Reconstitution of mGluR6-Gαo signaling in Xenopus oocytes indicates that RGS11·Gβ5-mediated GTPase acceleration in this system requires co-expression of R9AP. The results provide new insight into the regulation of mGluR6-Gαo signaling by the RGS11·Gβ5·R9AP complex and establish R9AP as a general GTPase-accelerating protein activity regulator of R7 RGS complexes. 相似文献