共查询到20条相似文献,搜索用时 0 毫秒
1.
Sequence and expression of a frog brain complementary DNA encoding a kainate-binding protein 总被引:11,自引:0,他引:11
K Wada C J Dechesne S Shimasaki R G King K Kusano A Buonanno D R Hampson C Banner R J Wenthold Y Nakatani 《Nature》1989,342(6250):684-689
Excitatory amino acids (EAAs) are important neurotransmitters in the vertebrate central nervous system. Electrophysiological and ligand-binding studies indicate that at least three different receptor subtypes for EAAs exist--N-methyl-D-aspartate, kainate and quisqualate receptor subtypes--on the basis of the preferred agonist of the receptors. We recently purified a kainate-binding protein (KBP) from frog (Rana pipiens berlandieri) brain by domoic acid (a high-affinity kainate analogue) affinity chromatography, and showed that the kainate-binding activity was associated with a protein of relative molecular mass 48,000 (Mr 48 K). The pharmacological properties and the anatomical distribution of KBP were consistent with those of a kainate receptor-ionophore complex. We have now isolated a complementary DNA encoding KBP of Mr 48 K. The deduced amino-acid sequence of the KBP has similar hydrophobic profiles to those found in other ligand-gated ion channel subunits, and shows some amino-acid sequence similarities to the corresponding regions of brain nicotinic acetylcholine receptor subunits. Localization of the KBP messenger RNAs by in situ hybridization histochemistry is compatible with the results of immunohistochemistry and receptor autoradiography studies. COS-7 cells transfected with the cDNA encoding the KBP show high-affinity kainate-binding activity with pharmacological properties similar to those of the biochemically purified KBP. These results provide the first molecular characterization of an EAA-binding site and raise the possibility that the KBP cDNA encodes a ligand-binding subunit of a kainate receptor-ionophore complex. 相似文献
2.
Primary structure and functional expression from complementary DNA of a brain calcium channel. 总被引:43,自引:0,他引:43
Y Mori T Friedrich M S Kim A Mikami J Nakai P Ruth E Bosse F Hofmann V Flockerzi T Furuichi 《Nature》1991,350(6317):398-402
The primary structure of a voltage-dependent calcium channel from rabbit brain has been deduced by cloning and sequencing the complementary DNA. Calcium channel activity expressed from the cDNA is dramatically increased by coexpression of the alpha 2 and beta subunits, known to be associated with the dihydropyridine receptor. This channel is a high voltage-activated calcium channel that is insensitive both to nifedipine and to omega-conotoxin. We suggest that it is expressed predominantly in cerebellar Purkinje cells and granule cells. 相似文献
3.
Cloning and expression of a complementary DNA encoding a bovine adrenal angiotensin II type-1 receptor 总被引:30,自引:0,他引:30
K Sasaki Y Yamano S Bardhan N Iwai J J Murray M Hasegawa Y Matsuda T Inagami 《Nature》1991,351(6323):230-233
Angiotensin II elicits different responses which affect cardiovascular, neuronal and electrolyte transport regulation. To understand the mechanisms responsible for its various actions, the receptor for angiotensin II has long been sought, but numerous attempts to purify the receptor have been unsuccessful owing to its instability and low concentration. We report here the expression cloning of a complementary DNA encoding a bovine angiotensin II receptor to overcome these difficulties. The receptor cDNA encodes a protein of 359 amino-acid residues with a transmembrane topology similar to that of other G protein-coupled receptors. COS-7 cells transfected with the cDNA expressed specific and high-affinity binding sites for angiotensin II, angiotensin II antagonist and a non-peptide specific antagonist for type-1 receptor. Dithiothreitol inhibited ligand binding. The concentration of intracellular Ca2+ and of inositol-1,4,5-trisphosphate increased in the transfected COS-7 cells in response to angiotensin II or angiotensin III, indicating that this receptor is the type-1 receptor for angiotensin II. Northern blot analysis revealed that the messenger RNA for this receptor is expressed in bovine adrenal medulla, cortex and kidney. 相似文献
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Primary structure and expression from complementary DNA of skeletal muscle ryanodine receptor 总被引:85,自引:0,他引:85
H Takeshima S Nishimura T Matsumoto H Ishida K Kangawa N Minamino H Matsuo M Ueda M Hanaoka T Hirose 《Nature》1989,339(6224):439-445
The sequence of 5,037 amino acids composing the ryanodine receptor from rabbit skeletal muscle sarcoplasmic reticulum has been deduced by cloning and sequencing the complementary DNA. The predicted structure suggests that the calcium release channel activity resides in the C-terminal region of the receptor molecule, whereas the remaining portion constitutes the 'foot' structure spanning the junctional gap between the sarcoplasmic reticulum and the transverse tubule. 相似文献
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Cloning, sequencing and expression of complementary DNA encoding the muscarinic acetylcholine receptor 总被引:75,自引:0,他引:75
T Kubo K Fukuda A Mikami A Maeda H Takahashi M Mishina T Haga K Haga A Ichiyama K Kangawa 《Nature》1986,323(6087):411-416
Cloning and sequence analysis of DNA complementary to porcine cerebral messenger RNA encoding the muscarinic acetylcholine receptor predict the complete amino-acid sequence of this protein. Expression of the complementary DNA produced functional muscarinic receptor in Xenopus oocytes. The muscarinic receptor is homologous with the beta-adrenergic receptor and rhodopsin in both amino-acid sequence and suggested transmembrane topography. 相似文献
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Cloning of a complementary DNA for a protein-tyrosine kinase that specifically phosphorylates a negative regulatory site of p60c-src. 总被引:55,自引:0,他引:55
The protein-tyrosine kinase activity of the proto-oncogene product p60c-src is negatively regulated by the phosphorylation of a tyrosine residue close to the C terminus, tyrosine 527. The phosphorylation might be catalysed by a so-far-unidentified tyrosine kinase, distinct from p60c-src. Recently we purified a protein-tyrosine kinase that specifically phosphorylates tyrosine 527 of p60c-src from neonatal rat brain. We have now confirmed the specificity of this enzyme by using a mutant p60c-src that has a phenylalanine instead of tyrosine 527, and cloned a complementary DNA that encodes the enzyme. The enzyme is similar to kinases of the src family in that it has two conserved regions, Src-homology regions 2 and 3, upstream of a tyrosine kinase domain. The amino-acid identity of each region is no more than 47%, however, and the enzyme lacks phosphorylation sites corresponding to tyrosines 416 and 527 of p60c-src and has no myristylation signal. These results suggest that this protein-tyrosine kinase, which might negatively regulate p60c-src, represents a new type of tyrosine kinase. 相似文献
10.
Human transforming growth factor-beta complementary DNA sequence and expression in normal and transformed cells 总被引:17,自引:0,他引:17
R Derynck J A Jarrett E Y Chen D H Eaton J R Bell R K Assoian A B Roberts M B Sporn D V Goeddel 《Nature》1985,316(6030):701-705
The partial amino-acid sequence of purified human transforming growth factor-beta (TGF-beta) was used to identify a series of cDNA clones encoding the protein. The cDNA sequence indicates that the 112-amino acid monomeric form of the natural TGF-beta homodimer is derived proteolytically from a much longer precursor polypeptide which may be secreted. TGF-beta messenger RNA is synthesized in various normal and transformed cells. 相似文献
11.
Isolation of complementary DNA clones encoding the human lymphocyte glycoprotein T1/Leu-1 总被引:4,自引:0,他引:4
N H Jones M L Clabby D P Dialynas H J Huang L A Herzenberg J L Strominger 《Nature》1986,323(6086):346-349
The T1/Leu-1/CD5 molecule, a human T-cell surface glycoprotein of relative molecular mass (Mr) 67,000, has been implicated in the proliferative response of activated T cells and in T-cell helper function. A similar involvement in T-cell proliferation has been reported for Ly-1, the murine homologue of T1. Here we report the complete amino-acid sequence of the T1 precursor molecule deduced from complementary DNA clones. The protein contains a classical signal peptide; a 347-amino-acid extracellular segment; a transmembrane region; and a 93-amino-acid intracellular segment. The extracellular segment contains many cysteine residues and is composed of two related structural domains separated by a proline/threonine-rich region. The T1 molecule has structural features characteristic of other receptor molecules. 相似文献
12.
Primary structure and functional expression from complementary DNA of the rod photoreceptor cyclic GMP-gated channel 总被引:35,自引:0,他引:35
U B Kaupp T Niidome T Tanabe S Terada W B?nigk W Stühmer N J Cook K Kangawa H Matsuo T Hirose 《Nature》1989,342(6251):762-766
The complete amino-acid sequence of the cyclic GMP-gated channel from bovine retinal rod photoreceptors, deduced by cloning and sequencing its complementary DNA, shows that the protein contains several putative transmembrane segments, followed by a region that is similar to the cyclic GMP-binding domains of cyclic GMP-dependent protein kinase. Expression of the complementary DNA produces cyclic GMP-gated channel activity in Xenopus oocytes. 相似文献
13.
Macrophages, granulocytes and many lymphocytes express or secrete receptors for the Fc domain of immunoglobulins (Ig). These Fc receptors (FcRs) are heterogeneous and can be distinguished on the basis of their cellular distribution and specificities for different immunoglobulin isotypes. Although their functions are not completely understood, FcRs are known to be involved in triggering various effector cell functions and in regulating differentiation and development of B-cells. One of the best characterized is the mouse macrophage-lymphocyte receptor for IgG1 and IgG2b (ref. 5). On macrophages, this FcR mediates the endocytosis of antibody-antigen complexes via coated pits and coated vesicles, the phagocytosis of Ig-coated particles, and the release of various inflammatory and cytotoxic agents. It is possible that the receptor possesses an intrinsic ligand-activated ion channel activity responsible for some of these functions. The IgG1/IgG2b FcR has been isolated and shown to be a transmembrane glycoprotein of relative molecular mass (Mr) 47,000-60,000 (47-60 K) containing four N-linked oligosaccharide chains and a large (greater than 10K) cytoplasmic domain. It is also immunologically indistinguishable from the murine Ly-17 alloantigen which, in turn, is tightly linked to the Mls lymphocyte activation locus. Here we describe the isolation and characterisation of a complementary DNA clone encoding the whole of the IgG1/IgG2b FcR expressed by the mouse macrophage-like cell line P388D1. The receptor is a member of the immunoglobulin superfamily and, like Ly-17, maps to the distal portion of chromosome 1. cDNA probes detect one or two mRNA species in FcR+ macrophage and B-cell lines, but not in FcR- cells or a receptor-deficient variant derived from a FcR+ B-cell line. Finally, DNA hybridization analysis indicates the receptor gene is partially deleted or rearranged in the FcR- variant. 相似文献
14.
Study of the mechanisms by which interferon (IFN) treatment of cells induces resistance to virus infections has been complicated by the multiple biochemical changes induced. Over 20 proteins are increased by IFN, including the double-stranded (ds) RNA-activated protein kinase, (2'-5') oligo A synthetase, surface proteins such as the major histocompatibility complex (MHC) proteins, and various proteins with unknown functions. The availability of cloned complementary DNAs for several IFN-induced proteins now allows us to probe their roles in IFN action. For instance, the murine Mx protein has been shown to confer resistance, to influenza virus. We studied chinese hamster ovary (CHO) cell clones expressing high constitutive levels of (2'-5') A synthetase as a result of transfection with the cDNA encoding the enzyme form which has a relative molecular mass (Mr) of 40K. Elevated enzyme correlates directly with resistance to infection by a picornavirus such as Mengo, but does not make the cells resistant to vesicular stomatitis virus (VSV). 相似文献
15.
Homology between P-glycoprotein and a bacterial haemolysin transport protein suggests a model for multidrug resistance 总被引:58,自引:0,他引:58
J H Gerlach J A Endicott P F Juranka G Henderson F Sarangi K L Deuchars V Ling 《Nature》1986,324(6096):485-489
Increased expression of P-glycoprotein, a plasma membrane glycoprotein of relative molecular mass (Mr) 170,000 (170K), occurs in a wide variety of cell lines that exhibit pleiotropic resistance to unrelated drugs. The presence of P-glycoprotein in human cancers refractory to chemotherapy suggests that tumour cells with multidrug resistance can arise during malignant progression. We have discovered striking homology between P-glycoprotein and the HlyB protein, a 66K Escherichia coli membrane protein required for the export of haemolysin (protein of Mr 107K). P-glycoprotein can be viewed as a tandem duplication of the HlyB protein. The hydropathy profiles of the two proteins are similar and reveal an extensive transmembrane region resembling those found in pore-forming plasma membrane proteins. The C-terminal region of P-glycoprotein and the HlyB protein contain sequences homologous to the nucleotide-binding domains of a group of closely related bacterial ATP-binding proteins. We propose a model for multidrug resistance in which P-glycoprotein functions as an energy-dependent export pump to reduce intracellular levels of anticancer drugs. 相似文献
16.
SunHui HuangQiman SuJin 《高技术通讯(英文版)》2005,11(1):75-79
Glutamine synthetase (GS, E.C.6.3.1.2) is a key enzyme involved in the assimilation of inorganic nitrogen in higher plants and gram-negative microorganisms. GS is the targeting enzyme of a herbicide phosphinothricin (PPT) or Basta. In order to generate PPT-resistant transgenic rice via overexpression of GS, we constructed a plant expression vector p2GS harboring two different isoenzymes GS1 and GS2 cDNAs under the control of constitutive promoters of rice Act1 and maize Ubiquitin(Ubi) genes. The p2GS was introduced into rice genome by Agrobacterium-mediated transformation and confirmed by PCR and Southern blot hybridization. GS-transgene expression was first detected by Northern blot analyses. Results from Basta test indicated that GS-transgenic plants can tolerate as high as 0.3% Basta solution. In addition, our results also demonstrated that GS overexpression conferred transformed rice calli PPT resistance. Thus, GS cassette can serve as a selective marker gene instead of bar cassette for selection of PPT transformants. 相似文献
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Isolation and ectopic expression of a bamboo MADS-box gene 总被引:1,自引:0,他引:1
TIANBo CHENYongyan YANYuanxin LIDezhu 《科学通报(英文版)》2005,50(3):217-224
A cDNA named DIMADS18 was isolated from the young spikelets of the sweet bamboo, Dendrocalamus latiflorus by RACE. DNA sequence analysis showed that DIMADS18 was composed of full ORF and 3UTR, but without 5UTR. The cDNA contained 1039 nucleotides and encoded a putative protein of 249 amino acid residues. The gene displayed the structure of a typical plant MADS box gene, which consisted of an MADS domain, K domain, a short I region, and the C-terminal region. Phylogenetic analysis of plant MADS box genes based on amino acid sequences revealed that DlMADS18 was grouped into the AGAMOUS-LIKE 6 (AGL6)-like subfamily. It was most likely homologous to the OsMADS6 of rice (Oryza sativa), with 88% sequence identity for the entire amino acid sequences. The DlMADS18 also showed relatively high amino acid sequence identity (59%) to AGL6 ofArabidopsis thaliana. To study the functions of DlMADS18, DlMADS18 cDNA clone driven by the CaMV 35S promoter was transformed into Arabidopsis plants. Transgenic plants of DlMADS18 exhibited the phenotypes of curled leaves, dwarfism, and early flowering with clustered terminal flowers. These results indicated that DlMADS18 may probably be involved in controlling the flowering time of D.latiflorus. 相似文献
19.
The yeast STE6 gene encodes a homologue of the mammalian multidrug resistance P-glycoprotein 总被引:82,自引:0,他引:82
Mammalian tumours displaying multidrug resistance overexpress a plasma membrane protein (P-glycoprotein), which is encoded by the MDR1 gene and apparently functions as an energy-dependent drug efflux pump. Tissue-specific expression of MDR1 and other members of the MDR gene family has been observed in normal cells, suggesting a role for P-glycoproteins in secretion. We have isolated a gene from the yeast Saccharomyces cerevisiae that encodes a protein very similar to mammalian P-glycoproteins. Deletion of this gene resulted in sterility of MATa, but not of MAT alpha cells. Subsequent analysis revealed that the yeast P-glycoprotein is the product of the STE6 gene, a locus previously shown to be required in MATa cells for production of a-factor pheromone. Our findings suggest that the STE6 protein functions to export the hydrophobic a-factor lipopeptide in a manner analogous to the efflux of hydrophobic cytotoxic drugs catalysed by the related mammalian P-glycoprotein. Thus, the evolutionarily conserved family of MDR-like genes, including the hlyB gene of Escherichia coli and the STE6 gene of S. cerevisiae, encodes components of secretory pathways distinct from the classical, signal sequence-dependent protein translocation system. 相似文献
20.
棉花细胞壁蛋白基因分离鉴定与表达分析 总被引:1,自引:0,他引:1
棉纤维起源于棉花胚珠外层表皮细胞.研究棉纤维发育,具有重要理论和实践意义.从棉纤维等组织cDNA文库中分离了186个棉花细胞壁结构蛋白基因,按照所编码的蛋白质结构特点,可将这些基因分为三类;富含脯氨酸细胞壁蛋白(Proline—rich cell wall proteins,PRPs)基因、伸展蛋白(Extensins)或者富含羟脯氨酸糖蛋白(Hydroxyproline—rich glycoprotein,HRGP)基因,富含甘氨酸蛋白(Glycine—rich proteins,GRPs)基因.采用cDNA microarray技术,比较上述基因在开花后10天的野生型棉花纤维和无絮无绒(fuzzless—lintless,fl)突变体胚珠表皮中表达谱发现,其中有7个基因在野生型纤维中特异性或高效性表达,表明它们可能与纤维发育有关. 相似文献