共查询到20条相似文献,搜索用时 0 毫秒
1.
Co-purification of Pea and Bean Leaf Soluble Auxin-binding Proteins with Ribulose-1,5-Bisphosphate Carboxylase 总被引:3,自引:3,他引:0
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Soluble auxin-binding proteins (ABPs) were purified to constant specific activity from bean and pea leaves by a procedure involving (NH4)2SO4 fractionation, anion exchange chromatography and gel filtration. Pea and bean ABPs exactly co-purify with ribulose-1,5-bisphosphate carboxylase (RuBPCase) in a variety of chromatographic separation procedures. The subunit compositions, electrophoretic purities and indole-3-acetic acid (IAA)-binding stoichiometries of the purified ABPs provide further evidence for the identity of RuBPCase and ABP. Pea ABP and bean ABP have dissociation constants for IAA of 0.8 and 1.3 micromolar, respectively, as determined by an (NH4)2SO4 precipitation assay for IAA-binding to insolubilized ABP. IAA can bind to soluble bean and pea ABP (RuBPCase) as determined by equilibrium dialysis with affinities and stoichiometries similar to those determined for insolubilized ABP. 相似文献
2.
A soluble cytokinin-binding protein from wheat germ that has a high affinity for a range of purine cytokinins also interacts with a variety of nonpurine compounds that can affect cytokinin-modified processes in animal or plant cells or which bind to proteins known to interact with certain cytokinins. A variety of structurally disparate compounds which inhibit chloroplast photosystem II activity (including phenylurea, carbanilate, and alkylamino-2-chloro-sym-triazine compounds) displace kinetin from the protein in an apparently competitive fashion. However, various energy transfer inhibitors (including organotin compounds and N,N′-dicy-clohexylcarbodiimide) also inhibit kinetin binding to the protein. N6,2-0′-Dibutyryl-3′,5′-cyclic AMP and 1-methyl-3-isobutylxanthine (the effects of which on fibroblast morphology and motility can be mimicked by cytokinins) are inhibitors of kinetin binding to the protein. A variety of compounds that can have antimitotic effects (including 1-methyl-3-isobutylxanthine and certain alkylated cyclic nucleotide, carbanilate, and tryptamine compounds) displace kinetin from the protein. However, a variety of indole derivatives also displace kinetin from the cytokinin-binding protein, which in a qualitative sense has a broad ligand specificity. 相似文献
3.
Shingo Sakai 《Bioscience, biotechnology, and biochemistry》2013,77(1):257-259
New acetylenic nematicidal compound, penipratynolene (1), methy (2′R)-4-(2′-hydroxy-3′-butynoxy)benzoate, together with two known compounds, 6-methoxycarbonylpicolinic acid (2) and 2,6-pyridinedicarboxylic acid (3), were isolated from the culture filtrate of Penicillium bilaiae Chalabuda. The structures of 1–3 were established by spectroscopic methods. The absolute configuration of 1 was confirmed by using a modified version of Mosher’s method. Compounds 1–3 showed nematicidal activity of 77%, 52%, and 98%, respectively, by a bioassay at 300 mg/l with the root-lesion nematode Pratylenchus penetrans. 相似文献
4.
5.
《Bioscience, biotechnology, and biochemistry》2013,77(3):504-507
A cytokinin-binding protein (CBP) was purified from a crude extract of etiolated mung bean seedlings by a protocol involving affinity chromatography on benzyladenine-linked Sepharose 4B, ion exchange chromatography on DEAE-Sephadex A50, and gel filtration on Sphacryl S-400. The molecular weight was estimatd to be about 200,000 by gel filtration. CBP appeared as two bands corresponding to molecular weights of about 45,000 and 48,000 on SDS-polyacrylamide gel electrophoresis. The dissociation constant for benzyladenine was 7.5 x 10-7 M. 14C-Benzyladenine-binding to CBP was reversible and could be inhibited by the addition of kinetin or trans-zeatin. Adenine, AMP, and ADP had no inhibitory effect on the binding of 14C-benzyladenine to CBP but the addition of ATP to the assay mixture enhanced the binding. 相似文献
6.
Nakajima Masatoshi; Sakai Shingo; Kanazawa Kenji; Kizawa Satoru; Yamaguchi Isomaro; Takahashi Nobutaka; Murofushi Noboru 《Plant & cell physiology》1993,34(2):289-296
A soluble binding protein specific for GA4, GA7 and GA9 waspartially purified from mung bean hypocotyls, and its characteristicswere examined. Affinity chromatography using immobilized GA3coupled to Sepharose 4B via the C-7 carboxyl group was veryeffective for purification of the protein. The molecular weightof the protein in its native state was estimated to be 150200kDa by gel-permeation chromatography. This protein may be aheterooligomer consisting of two subunits (23 kDa and 35 kDa).The optimum pH for binding of GA4 to the protein was around6.0 and the apparent dissociation constant (Kd) was 310-7 M. (Received April 24, 1992; Accepted December 16, 1992) 相似文献
7.
Brian T. Burmeister Li Wang Matthew G. Gold Randal A. Skidgel John P. O'Bryan Graeme K. Carnegie 《The Journal of biological chemistry》2015,290(19):12058-12067
Pathological cardiac hypertrophy (an increase in cardiac mass resulting from stress-induced cardiac myocyte growth) is a major factor underlying heart failure. Src homology 2 domain-containing phosphatase (Shp2) is critical for cardiac function because mutations resulting in loss of Shp2 catalytic activity are associated with congenital cardiac defects and hypertrophy. We identified a novel mechanism of Shp2 inhibition that may promote cardiac hypertrophy. We demonstrate that Shp2 is a component of the protein kinase A anchoring protein (AKAP)-Lbc complex. AKAP-Lbc facilitates PKA phosphorylation of Shp2, which inhibits Shp2 phosphatase activity. We identified two key amino acids in Shp2 that are phosphorylated by PKA. Thr-73 contributes a helix cap to helix αB within the N-terminal SH2 domain of Shp2, whereas Ser-189 occupies an equivalent position within the C-terminal SH2 domain. Utilizing double mutant PKA phosphodeficient (T73A/S189A) and phosphomimetic (T73D/S189D) constructs, in vitro binding assays, and phosphatase activity assays, we demonstrate that phosphorylation of these residues disrupts Shp2 interaction with tyrosine-phosphorylated ligands and inhibits its protein-tyrosine phosphatase activity. Overall, our data indicate that AKAP-Lbc integrates PKA and Shp2 signaling in the heart and that AKAP-Lbc-associated Shp2 activity is reduced in hypertrophic hearts in response to chronic β-adrenergic stimulation and PKA activation. Therefore, although induction of cardiac hypertrophy is a multifaceted process, inhibition of Shp2 activity through AKAP-Lbc-anchored PKA is a previously unrecognized mechanism that may promote this compensatory response. 相似文献
8.
生长素结合蛋白cDNA的克隆及其在黄瓜中的表达 总被引:6,自引:0,他引:6
利用RT_PCR方法从拟南芥 (Arabidopsisthaliana (L .)Heynh .)中克隆了生长素结合蛋白 (auxinbindingprotein 1 )cDNA ,进行了全序列测定。将该基因克隆在pBI1 2 1的 35S启动子和Nos终止子之间 ,得到植物表达载体p35EZ。通过根癌农杆菌 (Agrobacteriumtumefaciens (SmithetTownsend)Conn)介导的方法转化黄瓜 (CucumissativusL .)。对转基因植株进行了PCR和Southern检测。所得到的转基因黄瓜植株单性结实能力增强。 相似文献
9.
Specificity of Auxin-binding Sites on Maize Coleoptile Membranes as Possible Receptor Sites for Auxin Action 总被引:20,自引:12,他引:8
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Ray PM 《Plant physiology》1977,60(4):585-591
Dissociation coefficients of auxin-binding sites on maize (Zea mays L.) coleoptile membranes were measured, for 48 auxins and related ring compounds, by competitive displacement of 14C-naphthaleneacetic acid from the binding sites. The sites bind with high affinity several ring compounds with acidic side chains 2 to 4 carbons long, and much more weakly bind neutral ring compounds and phenols related to these active acids, most phenoxyalkylcarboxylic acids, and arylcarboxylic acids except benzoic acid, which scarcely binds, and triiodobenzoic acids, which bind strongly. Specificity of the binding is narrowed in the presence of a low molecular weight “supernatant factor” that occurs in maize and other tissues. Activity of many of the analogs as auxin agonists or antagonists in the cell elongation response was determined with maize coleoptiles. These activities on the whole roughly parallel the affinities of the binding sites for the same compounds, especially affinities measured in the presence of supernatant factor, but there are some quantitative discrepancies, especially among phenoxyalkylcarboxylic acids. In view of several factors that can cause receptor affinity and biological activity values to diverge quantitatively among analogs, the findings appear to support the presumption that the auxin-binding sites may be receptors for auxin action. 相似文献
10.
以黄瓜子房 (幼果 )RNA为模板 ,应用逆转录 聚合酶链式反应 (RT PCR) ,首次扩增出黄瓜生长素结合蛋白基因 (ABP1)cDNA片段 ,并进行测序和同源性分析。对ABP1基因在黄瓜子房 (幼果 )中的mRNA表达水平作了初步探讨 ,结果表明 ,该基因在开花前 1d的子房中表达信号较弱 ,在授粉后 2、4和 6d的幼果中表达增强 ;开花后 2d未经授粉的子房中 ,绿而膨大、能形成单性结实果者信号较强 ,黄而萎蔫、不能形成果实者信号较弱。Southern杂交结果表明 ,黄瓜生长素结合蛋白为小基因家族编码 相似文献
11.
S. K. Sathe 《Critical reviews in biotechnology》2013,33(2):175-223
ABSTRACT:?Dry beans are an important source of proteins, carbohydrates, dietary fiber, and certain minerals and vitamins in the human food supply. Among dry beans, Phaseolus beans are cultivated and consumed in the greatest quantity on a worldwide basis. Typically, most dry beans contain 15 to 25% protein on a dry weight basis (dwb). Water-soluble albumins and salt-soluble globulins, respectively, account for up to 10 to 30% and 45 to 70% of the total proteins (dwb). Dry bean albumins are typically composed of several different proteins, including lectins and enzyme inhibitors. A single 7S globulin dominates dry bean salt soluble fraction (globulins) and may account for up to 50 to 55% of the total proteins in the dry beans (dwb). Most dry bean proteins are deficient in sulfur amino acids, methionine, and cysteine, and therefore are of lower nutritional quality when compared with the animal proteins. Despite this limitation, dry beans make a significant contribution to the human dietary protein intake. In bean-based foods, dry bean proteins also serve additional functions that may include surface activity, hydration, and hydration-related properties, structure, and certain organoleptic properties. This article is intended to provide an overview of dry bean protein functionality with emphases on nutritional quality and hydration-related properties. 相似文献
12.
13.
Bean Leaf Expansion in Relation to Temperature 总被引:1,自引:0,他引:1
When dwarf Phaseolus vulgaris plants were grown in a controlledenvironment at 20, 25, 30, and 35° C, expansion of the primaryleaves occurred in two phases with an intermediate lag. Varyingrates and duration of expansion were involved, leading to greatestfinal areas at the two intermediate temperatures. Dry weightsof the leaves and leaf areas were similary influenced by temperature,except that the initial rates of increase continued for a longerperiod for weights than for areas. The rates of cell divisionand final numbers of cells were similar from 25 to 35° C,but both were decreased at 20° C. Final cell sizes were,on the other hand, decreased only at the highest temperature.The time trends of cell expansion varied greatly with temperature. Leaf expansion is discussed as a possible consequence of substratesupply, which may be determined by temperature in a number ofways. Cell division and cell expansion are not considered tobe joint direct determinants of leaf expansion. Temperatureinfluences division, with two consequences; the rate interactswith substrate supply to determine size of cells, and finalcell number affects potential leaf area. Cell size is regardedas being secondary to numbers of cells and total material available,although some factors can vary cell size independently of substrate,e.g. water status. An important control of leaf growth, until the attainment ofabout half the final area, may be exercised by way of the leaf.Subsequently, intra-plant competition is likely to dominate. 相似文献
14.
15.
16.
The transport of [14C]glycyl-glycine (Gly-Gly) has been characterized in leaf discs from mature exporting leaves of broad bean (Vicia faba L.). In terms of glycine (Gly) equivalents, the rate of transport of Gly-Gly was similar to that of Gly uptake. Uptake of Gly-Gly was localized mainly in the mesophyll cells, with little accumulation in the veins. It was optimal at pH 6.0, sensitive to thiol reagents and metabolic inhibitors, and exhibited a single saturable phase with an apparent Michaelis constant of 16 mM. Gly-Gly did not inhibit the uptake of labeled Gly. Addition of Gly-Gly induced a concentration-dependent pH rise in the medium, showing that peptide uptake is mediated with proton co-transport. Gly-Gly also induced a concentration-dependent transmembrane depolarization of mesophyll cells with an apparent Michaelis constant of 15 mM. This depolarization was followed by a transient hyperpolarization. When present at a 10-fold excess, various peptides and tripeptides were able to inhibit Gly-Gly uptake with the following decreasing order of efficiency: Gly-Gly-Gly = leucine-Gly > Gly-tyrosine > Gly-glutamine = Gly-glutamic acid > Gly-phenylalanine > Gly-threonine > Gly-aspartic acid = Gly-asparagine = aspartic acid-Gly. Gly inhibited the uptake of Gly-Gly only slightly, whereas tetraGly and the tripeptide glutathione were not inhibitory. The dipeptides inhibiting Gly-Gly uptake also induced changes in the transmembrane potential difference of mesophyll cells and were able to affect in a complex way the response normally induced by Gly-Gly. Altogether, the data demonstrate the existence of a low-affinity, broad-specificity H+/peptide co-transporter at the plasma membrane of mesophyll cells. The physiological importance of this transporter for the exchange of nitrogenous compounds in mature leaves remains to be determined, as do the details of the electrophysiological events induced by the dipeptides. 相似文献
17.
James J. Havranek 《The Journal of biological chemistry》2010,285(41):31095-31099
A long-standing goal of computational protein design is to create proteins similar to those found in Nature. One motivation is to harness the exquisite functional capabilities of proteins for our own purposes. The extent of similarity between designed and natural proteins also reports on how faithfully our models represent the selective pressures that determine protein sequences. As the field of protein design shifts emphasis from reproducing native-like protein structure to function, it has become important that these models treat the notion of specificity in molecular interactions. Although specificity may, in some cases, be achieved by optimization of a desired protein in isolation, methods have been developed to address directly the desire for proteins that exhibit specific functions and interactions. 相似文献
18.
19.
Hye Jin Kang Angela D. Wilkins Olivier Lichtarge Theodore G. Wensel 《The Journal of biological chemistry》2015,290(5):2870-2878
To determine the structural origins of diverse ligand response specificities among metabotropic glutamate receptors (mGluRs), we combined computational approaches with mutagenesis and ligand response assays to identify specificity-determining residues in the group I receptor, mGluR1, and the group III receptors, mGluR4 and mGluR7. Among these, mGluR1 responds to l-glutamate effectively, whereas it binds weakly to another endogenous ligand, l-serine-O-phosphate (l-SOP), which antagonizes the effects of l-glutamate. In contrast, mGluR4 has in common with other group III mGluR that it is activated with higher potency and efficacy by l-SOP. mGluR7 differs from mGluR4 and other group III mGluR in that l-glutamate and l-SOP activate it with low potency and efficacy. Enhanced versions of the evolutionary trace (ET) algorithm were used to identify residues that when swapped between mGluR1 and mGluR4 increased the potency of l-SOP inhibition relative to the potency of l-glutamate activation in mGluR1 mutants and others that diminished the potency/efficacy of l-SOP for mGluR4 mutants. In addition, combining ET identified swaps from mGluR4 with one identified by computational docking produced mGluR7 mutants that respond with dramatically enhanced potency/efficacy to l-SOP. These results reveal that an early functional divergence between group I/II and group III involved variation at positions primarily at allosteric sites located outside of binding pockets, whereas a later divergence within group III occurred through sequence variation both at the ligand-binding pocket and at loops near the dimerization interface and interlobe hinge region. They also demonstrate the power of ET for identifying allosteric determinants of evolutionary importance. 相似文献
20.
Properties of a Solubilized Microsomal Auxin-binding Protein from Coleoptiles and Primary Leaves of Zea mays 总被引:13,自引:10,他引:3
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
An auxin-binding protein can be solubilized from microsomal membranes of Zea mays using either Triton X-100 extraction of the membranes or buffer extraction of the acetone-precipitated membranes. This paper describes the properties of the binding protein solubilized by these two methods. The binding is assayed by gel filtration chromatography in the presence of naphthalene [2-14C]acetic acid. Binding is rapid and reversible with an optimum at pH 5. Both preparations show similar molecular weights by gel filtration (80,000 daltons) at pH 7.6 and 0.1 molar NaCl, and both aggregate at low ionic strength. They appear to be the same active molecular species. The binding activity is destroyed by trypsin, pronase or para-chloromercuribenzoic acid, but not significantly reduced by phospholipase C, DNase, RNase, or dithioerythritol. Since saturating amounts of naphthalene acetic acid protect the molecule from inhibition by para-chloromercuribenzoic acid, it is concluded that the binding protein has a sulfhydryl group at the binding site, or protects such a group in its binding conformation. The dissociation constant of the protein for naphthalene acetic acid is 4.6 × 10−8 molar with 30 picomoles of sites per gram of tissue fresh weight. Binding constants were estimated for 13 other natural and synthetic auxins by competition with naphthalene[2-14C]acetic acid. Their dissociation constants are in general agreement with published values for their binding to intact membranes and their biological activity, although several exceptions were noted. A supernatant factor from the same tissue changes the apparent affinity of the protein for naphthalene acetic acid. This factor may be the same one as has been previously reported to alter the affinity of intact microsomes for auxin. 相似文献