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1.
In the 60 S ribosomal subunit, the lateral stalk made of the P-proteins plays a major role in translation. It contains P0, an insoluble protein anchoring P1 and P2 to the ribosome. Here, rat recombinant P0 was overproduced in inclusion bodies and solubilized in complex with the other P-proteins. This method of solubilization appeared suitable to show protein complexes and revealed that P1, but not P2, interacted with P0. Furthermore, the use of truncated mutants of P1 and P2 indicated that residues 1-63 in P1 connected P0 to residues 1-65 in P2. Additional experiments resulted in the conclusion that P1 and P2 bound one another, either connected with P0 or free, as found in the cytoplasm. Accordingly, a model of association for the P-proteins in the stalk is proposed. Recombinant P0 in complex with phosphorylated P2 and either P1 or its (1-63) domain efficiently restored the proteosynthetic activity of 60 S subunits deprived of native P-proteins. Therefore, refolded P0 was functional and residues 1-63 only in P1 were essential. Furthermore, our results emphasize that the refolding principle used here is worth considering for solubilizing other insoluble proteins. 相似文献
2.
The aim was to investigate the roles of proline residues in extracellular loop 2 (P172, P183, P188 and P209) and transmembrane domains 2, 5, 11 and 12 (P108, P270, P526, P551, P552 and P570) in determining noradrenaline transporter (NET) expression and function. Mutants of human NET with these residues mutated to alanine were pharmacologically characterized. Mutation of P108, P270 and P526 disrupted cell surface expression, from [3H]nisoxetine binding and confocal microscopy data. Mutations of P526, P551 and P570 reduced transporter turnover (Vmax of [3H]noradrenaline uptake/Bmax of [3H]nisoxetine binding) by 1.5-1.7-fold compared with wild-type NET, so these residues might be involved in conformational changes associated with substrate translocation. Conversely, mutations of P172, P183, P188 and P209 increased Vmax/Bmax by 2-3-fold compared with wild-type, indicating that the presence of these proline residues limits turnover of the NET. The mutations had few effects on apparent affinities of substrates or affinities of inhibitors, except decreases in inhibitor affinities after mutations of the P270 and P570 residues, and increases after mutation of the P526 residue. Hence, proline residues in extracellular loop 2 and in transmembrane domains have a range of roles in determining expression and function of the NET. 相似文献
3.
Przyboś E 《Folia biologica》2005,53(1-2):61-63
Among 15 species of the Paramecium aurelia complex known world-wide, 10 have been found in Europe, namely: P. primaurelia, P. biaurelia, P. triaurelia, P. tetraurelia, P. pentaurelia, P. sexaurelia, P. septaurelia, P. novaurelia, P. dodecaurelia, and P. tredecaurelia. Recent data on the frequency of occurrence of the species in Europe are given in the paper. 相似文献
4.
Kentaro Baba Kazuhiro Tumuraya Isao Tanaka Min Yao Toshio Uchiumi 《Nucleic acids research》2013,41(6):3635-3643
In animal ribosomes, two stalk proteins P1 and P2 form a heterodimer, and the two dimers, with the anchor protein P0, constitute a pentameric complex crucial for recruitment of translational GTPase factors to the ribosome. To investigate the functional contribution of each copy of the stalk proteins, we constructed P0 mutants, in which one of the two C-terminal helices, namely helix I (N-terminal side) or helix II (C-terminal side) were unable to bind the P1–P2 dimer. We also constructed ‘one-C-terminal domain (CTD) stalk dimers’, P1–P2ΔC and P1ΔC–P2, composed of intact P1/P2 monomer and a CTD-truncated partner. Through combinations of P0 and P1–P2 variants, various complexes were reconstituted and their function tested in eEF-2-dependent GTPase and eEF-1α/eEF-2-dependent polyphenylalanine synthesis assays in vitro. Double/single-CTD dimers bound to helix I showed higher activity than that bound to helix II. Despite low polypeptide synthetic activity by a single one-CTD dimer, its binding to both helices considerably increased activity, suggesting that two stalk dimers cooperate, particularly in polypeptide synthesis. This promotion of activity by two stalk dimers was lost upon mutation of the conserved YPT sequence connecting the two helices of P0, suggesting a role for this sequence in cooperativity of two stalk dimers. 相似文献
5.
The lateral ribosomal stalk is responsible for the kingdom-specific binding of translation factors and activation of GTP hydrolysis during protein synthesis. The eukaryotic stalk is composed of three acidic ribosomal proteins P0, P1 and P2. P0 binds two copies of P1/P2 hetero-dimers to form a pentameric P-complex. The structure of the eukaryotic stalk is currently not known. To provide a better understanding on the structural organization of eukaryotic stalk, we have determined the solution structure of the N-terminal dimerization domain (NTD) of P1/P2 hetero-dimer. Helix-1, -2 and -4 from each of the NTD-P1 and NTD-P2 form the dimeric interface that buries 2200 A(2) of solvent accessible surface area. In contrast to the symmetric P2 homo-dimer, P1/P2 hetero-dimer is asymmetric. Three conserved hydrophobic residues on the surface of NTD-P1 are replaced by charged residues in NTD-P2. Moreover, NTD-P1 has an extra turn in helix-1, which forms extensive intermolecular interactions with helix-1 and -4 of NTD-P2. Truncation of this extra turn of P1 abolished the formation of P1/P2 hetero-dimer. Systematic truncation studies suggest that P0 contains two spine-helices that each binds one copy of P1/P2 hetero-dimer. Modeling studies suggest that a large hydrophobic cavity, which can accommodate the loop between the spine-helices of P0, can be found on NTD-P1 but not on NTD-P2 when the helix-4 adopts an 'open' conformation. Based on the asymmetric properties of NTD-P1/NTD-P2, a structural model of the eukaryotic P-complex with P2/P1:P1/P2 topology is proposed. 相似文献
6.
Synthetic peptides as model substrates for the study of the specificity of the polycation-stimulated protein phosphatases 总被引:3,自引:0,他引:3
P Agostinis J Goris L A Pinna F Marchiori J W Perich H E Meyer W Merlevede 《European journal of biochemistry》1990,189(2):235-241
The substrate specificity of the different forms of the polycation-stimulated (PCS, type 2A) protein phosphatases and of the active catalytic subunit of the ATP, Mg-dependent (type 1) phosphatase (AMDC) was investigated, using synthetic peptides phosphorylated by either cyclic-AMP-dependent protein kinase or by casein kinase-2. The PCS phosphatases are very efficient toward the Thr(P) peptides RRAT(P)VA and RRREEET(P)EEE when compared with the Ser(P) analogues RRAS(P)VA and RRREEES(P)EEEAA. Despite their distinct sequence, both Thr(P) peptides are excellent substrates for the PCSM and PCSH1 phosphatases, being dephosphorylated faster than phosphorylase a. The slow dephosphorylation of RRAS(P)VA by the PCS phosphatases could be increased substantially by the insertion of N-terminal (Arg) basic residues. In contrast with the latter, the AMDC phosphatase shows very poor activity toward all the phosphopeptides tested, without preference for either Ser(P) or Thr(P) peptides. However, N-terminal basic residues also favor the dephosphorylation of otherwise almost inert substrates by the AMDC phosphatase. Hence, while the dephosphorylation of Thr(P) substrates by the PCS phosphatases is highly favored by the nature of the phosphorylated amino acid, phosphatase activity toward Ser(P)-containing peptides may require specific determinants in the primary structure of the phosphorylation site. 相似文献
7.
Serological studies on the major histocompatibility complex of new inbred strains of the guinea pig.
New inbred strains of guinea pigs, JY 1, JY 2, JY 3, JY 6, JY 9 and JY 10 have been established in this Institute. Serologic studies of guinea pig leukocyte antigens (GPLA antigens) were carried out in order to examine their major histocompatibility complex (GPLA complex). Antisera specific for Ia antigens were raised by cross-immunization of NIH strain 2 (NIH 2) and NIH strain 13 (NIH 13) guinea pigs, well known inbred guinea pigs. The sera identified four distinct Ia specificities, which were designated as Ia.2a, Ia2b, Ia.13a and Ia.13b. Six antigenic specificities different from the above Ia specificities were identified by sera obtained by appropriate immunization of the inbred guinea pigs and were designated as P.1, P.2, P.3, P.4, P.5 and P.6. Antigenic specificities of GPLA antigens recognized in inbred guinea pigs were : NIH 2 (Ia.2a, Ia.2b, P.1, P.2, P.O, P.4), NIH 13 (Ia.13a, Ia.13b, P.1, P.2, P.3, P.4), JY 1 (Ia.13a, Ia.13b, P.5), JY 2 (Ia.2b, Ia.13b, P.3, P.4, P.6), JY 3 (Ia.13a, Ia.13b, P.2, P.4, P.5), JY 6 (Ia.2b, Ia.13b, P.3, P.6), JY 9 (Ia.13a, Ia.13b, P.4, P.5), JY 10 (Ia.13a, Ia.13b, P.2, P.3, P.4, P.6), JY 9 (Ia.13a, Ia.13b, P.4, P.5), JY 10 (Ia.13a, Ia.13b, P.2, P.3, P.4, P.6). The correspondence of these specificities to those already reported was discussed and the P.2 or P.4 was considered to be an additional specificity of GPLA antigens that have not been reported yet. As the new inbred strains of guinea pigs were thus found to possess characteristic GPLA complex, which differ from each other and from those of NIH 2 and NIH 13 strain, they should be useful for studies of roles of the major histocompatibility complex in the immune system. 相似文献
8.
This study contrasts the ovipositional profiles of four members of the Papilio glaucus group, P. glaucus , P. multicaudatus , P. canadensis , and P. rutulus. We used seven choice oviposition bioassays containing leaves from hosts in seven plant families utilized by members of the P. glaucus group. Specifically, we contrast the overall ovipositional profiles of these species and their acceptance of a host in a novel plant family ( Populus tremuloides : Salicaceae) and a host in a putatively ancestral host plant family ( Liriodendron tulipifera : Magnoliaceae). Significant differences were observed between the ovipositional profiles of P. glaucus and P. multicaudatus relative to each other and to P. canadensis and P. rutulus. In contrast, no significant differences were observed between the ovipositional profiles of P. canadensis and P. rutulus , which were also the only species that accepted P. tremuloides. Unlike the acceptance of P. tremuloides , the acceptance of L. tulipifera was present throughout the group despite the inability of the larvae of most species in the group to utilize this host. These results support the prediction of the "hierarchical threshold model" that ancestral host plants are likely to be retained in the ovipositional hierarchy while novel hosts should only be accepted by derived populations. 相似文献
9.
Ecke D Hanck T Tulapurkar ME Schäfer R Kassack M Stricker R Reiser G 《The Biochemical journal》2008,409(1):107-116
Nucleotides signal through purinergic receptors such as the P2 receptors, which are subdivided into the ionotropic P2X receptors and the metabotropic P2Y receptors. The diversity of functions within the purinergic receptor family is required for the tissue-specificity of nucleotide signalling. In the present study, hetero-oligomerization between two metabotropic P2Y receptor subtypes is established. These receptors, P2Y1 and P2Y11, were found to associate together when co-expressed in HEK293 cells. This association was detected by co-pull-down, immunoprecipitation and FRET (fluorescence resonance energy transfer) experiments. We found a striking functional consequence of the interaction between the P2Y11 receptor and the P2Y1 receptor where this interaction promotes agonist-induced internalization of the P2Y11 receptor. This is remarkable because the P2Y11 receptor by itself is not able to undergo endocytosis. Co-internalization of these receptors was also seen in 1321N1 astrocytoma cells co-expressing both P2Y11 and P2Y1 receptors, upon stimulation with ATP or the P2Y1 receptor-specific agonist 2-MeS-ADP. 1321N1 astrocytoma cells do not express endogenous P2Y receptors. Moreover, in HEK293 cells, the P2Y11 receptor was found to functionally associate with endogenous P2Y1 receptors. Treatment of HEK293 cells with siRNA (small interfering RNA) directed against the P2Y1 receptor diminished the agonist-induced endocytosis of the heterologously expressed GFP-P2Y11 receptor. Pharmacological characteristics of the P2Y11 receptor expressed in HEK293 cells were determined by recording Ca2+ responses after nucleotide stimulation. This analysis revealed a ligand specificity which was different from the agonist profile established in cells expressing the P2Y11 receptor as the only metabotropic nucleotide receptor. Thus the hetero-oligomerization of the P2Y1 and P2Y11 receptors allows novel functions of the P2Y11 receptor in response to extracellular nucleotides. 相似文献
10.
Characterization of the DNA-protein complex at the termini of the bacteriophage PRD1 genome. 总被引:24,自引:5,他引:19
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DNA of bacteriophage PRD1 has protein P8 at its termini. Extracts of infected cells are able to derivatize P8 in vitro with labeled dGTP. Two early proteins, P1 and P8, products of genes I and VIII, respectively, are the only phage proteins necessary for the formation of the protein P8-dGMP complex. This was shown by complementation of extracts from cells infected with mutants and by use of extracts from cells carrying cloned genes I and VIII. With Escherichia coli mutants that are temperature sensitive for DNA synthesis, it was possible to show that the formation of the protein P8-dGMP complex was dependent upon the host replication apparatus. The analysis of the purified protein P8-dGMP complex by hydrolysis and enzymatic digestion showed that there is a covalent phosphodiester bond between tyrosine and 5'-dGMP. 相似文献
11.
Koshimizu T Koshimizu M Stojilkovic SS 《The Journal of biological chemistry》1999,274(53):37651-37657
The P2X purinergic receptor channels (P2XRs) differ among themselves with respect to the rates of desensitization during prolonged agonist stimulation. Here we studied the desensitization of recombinant channels by monitoring the changes in intracellular free Ca(2+) concentration in cells stimulated with ATP, the native and common agonist for all P2XRs. The focus in our investigations was on the relevance of the P2XR C terminus in controlling receptor desensitization. When expressed in GT1 cells, the P2XRs desensitized with rates characteristic to each receptor subtype: P2X(1)R = P2X(3)R > P2X(2b)R > P2X(4)R > P2X(2a)R > P2X(7)R. A slow desensitizing pattern of P2X(2a)R was mimicked partially by P2X(3)R and fully by P2X(4)R when the six-amino acid sequences of these channels located in the cytoplasmic C terminus were substituted with the corresponding arginine 371 to proline 376 sequence of P2X(2a)R. Changing the total net charge in the six amino acids of P2X(4)R to a more positive direction also slowed the receptor desensitization. On the other hand, substitution of arginine 371-proline 376 sequence of P2X(2a)R with the corresponding sequences of P2X(1)R, P2X(3)R, and P2X(4)R increased the rate of receptor desensitization. Furthermore, heterologous polymerization of wild-type P2X(2a)R and mutant P2X(3)R having the C-terminal six amino acids of P2X(2a)R at its analogous position resulted in a functional channel whose desensitization was significantly delayed. These results suggest that composition of the C-terminal six-amino acid sequence and its electrostatic force influence the rate of receptor desensitization. 相似文献
12.
The RNA phosphodiester bond cleavage activity of a series of 16 thio-deoxyribozymes 10-23, containing a P-stereorandom single phosphorothioate linkage in predetermined positions of the catalytic core from P1 to P16, was evaluated under single-turnover conditions in the presence of either 3 mM Mg(2+) or 3 mM Mn(2+). A metal-specificity switch approach permitted the identification of nonbridging phosphate oxygens (proR(P) or proS(P)) located at seven positions of the core (P2, P4 and P9-13) involved in direct coordination with a divalent metal ion(s). By contrast, phosphorothioates at positions P3, P6, P7 and P14-16 displayed no functional relevance in the deoxyribozyme-mediated catalysis. Interestingly, phosphorothioate modifications at positions P1 or P8 enhanced the catalytic efficiency of the enzyme. Among the tested deoxyribozymes, thio-substitution at position P5 had the largest deleterious effect on the catalytic rate in the presence of Mg(2+), and this was reversed in the presence of Mn(2+). Further experiments with thio-deoxyribozymes of stereodefined P-chirality suggested direct involvement of both oxygens of the P5 phosphate and the proR(P) oxygen at P9 in the metal ion coordination. In addition, it was found that the oxygen atom at C6 of G(6) contributes to metal ion binding and that this interaction is essential for 10-23 deoxyribozyme catalytic activity. 相似文献
13.
Bacteriophage 6 is a complex enveloped double-stranded RNA virus with a segmented genome and replication strategy quite similar to that of the Reoviridae. An in vitro packaging and replication system using purified components is available. The positive-polarity genomic segments are translocated into a preformed polymerase complex (procapsid) particle. This particle is composed of four proteins: the shell-forming protein P1, the RNA polymerase P2, and two proteins active in packaging. Protein P7 is involved in stable packaging, and protein P4 is a homomultimeric potent nucleoside triphosphatase that provides the energy for the RNA translocation event. In this investigation, we used mutational analysis to study P4 multimerization and assembly. P4 is assembled onto a preformed particle containing proteins P2 and P7 in addition to P1. Only simultaneous production of P1 and P4 in the same cell leads to P4 assembly on P1 alone, whereas the P1 shell is incompetent for accepting P4 if produced separately. The C-terminal part of P4 is essential for particle assembly but not for multimerization or enzymatic activity. Altering the P4 nucleoside triphosphate binding site destroys the ability to form multimers. 相似文献
14.
Akio Yamazaki Masahiro Tatsumi Vladimir A. Bondarenko Sadamu Kurono Naoka Komori Hiroyuki Matsumoto Isao Matsuura Fumio Hayashi Russell K. Yamazaki Jiro Usukura 《Molecular and cellular biochemistry》2010,339(1-2):235-251
Rod photoreceptor cGMP phosphodiesterase (PDE6) consists of a catalytic subunit complex (Pαβ) and two inhibitory subunits (Pγ). In the accompanying article, using bovine photoreceptor outer segment homogenates, we show that Pγ as a complex with the GTP-bound transducin α subunit (GTP-Tα) dissociates from Pαβγγ on membranes, and the Pαβγγ becomes Pγ-depleted. Here, we identify and characterize the Pγ-depleted PDE. After incubation with or without guanosine 5′-O-(3-thiotriphosphate) (GTPγS), Pαβ complexes are extracted. When a hypotonic buffer is used, Pαβγγ, Pαβγ, and a negligible amount of a Pαβ complex containing Pγ are isolated with GTPγS, and only Pαβγγ is obtained without GTPγS. When an isotonic buffer containing Pδ, a prenyl-binding protein, is used, Pαβγγδ, Pαβγδδ, and a negligible amount of a Pαβ complex containing Pγ and Pδ are isolated with GTPγS, and Pαβγγδ is obtained without GTPγS. Neither Pαβ nor Pαβγγ complexed with GTPγS-Tα is found under any condition we examined. Pαβγ has ~12 times higher PDE activity and ~30 times higher Pγ sensitivity than those of Pαβγγ. These results indicate that the Pγ-depleted PDE is Pαβγ. Isolation of Pαβγγδ and Pαβγδδ suggests that one C-terminus of Pαβ is involved in the Pαβγγ interaction with membranes, and that Pγ dissociation opens another C-terminus for Pδ binding, which may lead to the expression of high PDE activity. Cone PDE behaves similarly to rod PDE in the anion exchange column chromatography. We conclude that the mechanisms for PDE activation are similar in mammalian and amphibian photoreceptors as well as in rods and cones. 相似文献
15.
KNUT ROGNES 《Systematic Entomology》1991,16(4):439-498
Abstract. The Pollenia viatica species-group ( =pallida species-group) is defined and described. It consists of P.bicolor Robineau-Desvoidy, P.bulgarica Jacentkovsk, P.fulvipalpis Macquart, P.mediterranea Grunin, P.ruficrura Rondani, stat.rev., P.viatica Robineau-Desvoidy, stat.rev. (= P.pallida Rodendorf, syn.n.) and P.ponti sp.n. It is mono-phyletic, mainly on the basis of a synapomorphic ovipositor tip, and the vagabunda species-group is its sister group. A key to all species is provided, the terminalia of both sexes illustrated and all taxa redescribed. Lectotypes are designated for Pollenia bisulca Pandellé, P.ruficrura Rondani and P.viatica Robineau-Desvoidy. Pollenia ruficrura is removed from its current status as a nomen dubium. Pollenia bicolor and P.ponti are western Mediterranean species, the latter also occurs in Czechoslovakia. P.fulvipalpis is endemic to France and Channel Islands. P.ruficrura is known only from Corsica and Italy. P.mediterranea is an eastern Mediterranean species reaching east to Armenia. P.bulgarica is distributed from Hungary to Caucasus. P.viatica is widespread from France, Great Britain and Southern Scandinavia to Central Asia. All species are summer or autumn species, and seem to have only one generation each year. The immature stages and life-cycle of a species from Paris, France, named 'Pollenia rudis' by Keilin (1909, 1915) are tentatively assigned to P. viatica . It survives the winter as a first instar larva within the earthworm Allolobophora chlorotica Savigny. 相似文献
16.
黄颡鱼属物种的RAPD分子鉴定及杂种遗传分析 总被引:10,自引:2,他引:10
使用20个随机引物研究了4种黄颡鱼的RAPD图谱,其中6个可以用来准确鉴别4个种,分别是S1、S2、S5、S7、S8、S17。同时也分析了4种黄颡鱼之间的亲缘关系,发现种间遗传距离D值在0.5000左右波动,黄颡鱼-叉尾黄颡鱼、瓦氏黄颡鱼-光泽黄颡鱼之间的D值最小,分别是0.4816和0.4017。聚类图显示,黄颡鱼和叉尾黄颡鱼属同一分支;瓦氏黄颡鱼和光泽黄颡鱼属另一分支,说明它们之间分别有更近的亲缘关系。另外,采用黄颡鱼作母本,瓦氏黄颡鱼作父本,获得了杂交F1代,对F1代进行RAPD分析,发现了杂种遗传图谱的三种变化情况:叠加、叠加-变异、叠加-弱化,表明F1代DNA多态性增强,杂合性提高,预示着杂种优势的可能性。
相似文献
17.
In this paper, the genusPinguicula is revised in the Iberian Peninsula. The taxa studied areP. vulgaris, P. nevadensis, P. grandiflora subsp.grandiflora, P. dertosensis, P. vallisneriifolia, P. longifolia subsp.longifolia, P. mundi, P. alpina andP. lusitanica. Five taxa are endemic to the area studied (P. nevadensis, P. dertosensis, P. vallisneriifolia, P. longifolia subsp.longifolia andP. mundi). Two of them are confined to restricted areas:P. nevadensis is endemic to the Sierra Nevada (S Spain) andP. longifolia subsp.longifolia is endemic to the central Pyrenees (N Spain). Taxonomical, nomenclatural, morphological, chromosomal, biogeographical and ecological data are reported for each taxon. A new combination is proposed:Pinguicula Sect.Longifolia (Casper) G. Blanca, M. Ruíz-Rejón etR. Zamora. 相似文献
18.
On the role of IS1 in the formation of hybrids between the bacteriophage P1 and the R plasmid NR1 总被引:13,自引:0,他引:13
Summary The genomes of bacteriophage P1 derivatives carrying drug resistance genes derived from an R plasmid NR1 were analysed by restriction cleavage and be DNA-DNA hybridization. Two representatives of a class of oversized P1CmSmSu phages were identified as P1 carrying the entire r-determinant of NR1 together with its two flanking, directly repeated IS1. In one case the r-determinant insertion is carried at the site of the residential IS1 of P1, in the other case it is transposed into another region of the P1 genome. Models postulate that the first type resulted from reciprocal recombination within IS1 elements and that the formation of the second type of P1-R hybrid depended both on IS1 mediated transposition and reciprocal recombination. Plaque forming P1Cm or P1CmSm phages are explained as IS1 mediated deletion derivatives of P1CmSmSu, although an alternative model postulates that sometimes P1Cm phages might result from two consecutive transposition events of only one IS1 without involving reciprocal recombination. Secondary P1 derivatives carrying only one IS1 at the site of the original r-determinant or of Cm insertions into P1 must have been produced by reciprocal recombination between the two IS1 flanking the insertions. An implication from this study, that any genetic material carried adjacent to an IS1 element may undergo passive transposition, is discussed. 相似文献
19.
Tchórzewski M Boldyreff B Issinger OG Grankowski N 《The international journal of biochemistry & cell biology》2000,32(7):737-746
The surface acidic ribosomal proteins (P-proteins), together with ribosomal core protein P0 form a multimeric lateral protuberance on the 60 S ribosomal subunit. This structure, also called stalk, is important for efficient translational activity of the ribosome. In order to shed more light on the function of these proteins, we are the first to have precisely analyzed mutual interactions among human P-proteins, employing the two hybrid system. The human acidic ribosomal P-proteins, (P1 or P2,) were fused to the GAL4 binding domain (BD) as well as the activation domain (AD), and analyzed in yeast cells. It is concluded that the heterodimeric complex of the P1/P2 proteins is formed preferentially. Formation of homodimers (P1/P1 and P2/P2) can also be observed, though with much less efficiency. Regarding that, we propose to describe the double heterodimeric complex as a protein configuration which forms the 60 S ribosomal stalk: P0-(P1/P2)(2). Additionally, mutual interactions among human and yeast P-proteins were analyzed. Heterodimer formation could be observed between human P2 and yeast P1 proteins. 相似文献
20.
W R Haines K Migita J A Cox T M Egan M M Voigt 《The Journal of biological chemistry》2001,276(35):32793-32798
Based on pharmacological properties, the P2X receptor family can be subdivided into those homo-oligomers that are sensitive to the ATP analog alphabeta-methylene ATP(alphabetameATP) (P2X(1) and P2X(3)) and those that are not (P2X(2), P2X(4), P2X(5), P2X(6), and P2X(7)). We exploited this dichotomy through the construction of chimeric receptors and site-directed mutagenesis in order to identify domains responsible for these differences in the abilities of extracellular agonists to gate P2X receptors. Replacement of the extracellular domain of the alphabetameATP-sensitive rat P2X(1) subunit with that of the alphabetameATP-insensitive rat P2X(2) subunit resulted in a receptor that was still alphabetameATP-sensitive, suggesting a non-extracellular domain was responsible for the differential gating of P2X receptors by various agonists. Replacement of the first transmembrane domain of the rat P2X(2) subunit with one from an alphabetameATP-sensitive subunit (either rat P2X(1) or P2X(3) subunit) converted the resulting chimera to alphabetameATP sensitivity. This conversion did not occur when the first transmembrane domain came from a non-alphabetameATP-sensitive subunit. Site-directed mutagenesis indicated that the C-terminal portion of the first transmembrane domain was important in determining the agonist selectivity of channel gating for these chimeras. These results suggest that the first transmembrane domain plays an important role in the agonist operation of the P2X receptor. 相似文献