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101.
Immunocytolocalization of glutamine synthetase in mesophyll and phloem of leaves ofSolanum tuberosum L. 总被引:1,自引:0,他引:1
Summary Localization of glutamine synthetase inSolanum tuberosum leaves was investigated by techniques of Western tissue printing and immunogold electron microscopy. Anti-GS antibodies used in immunolocalization recognize two peptides (45 kDa and 42 kDa) on Western blots. Antibody stained tissue prints on nitrocellulose membranes allowed low resolution localization of GS. Immunostaining was most evident in the adaxial phloem of the leaf midribs and petiole veins. High-resolution localization of glutamine synthetase by immunogold electron microscopy revealed that this enzyme occurs in both the chloroplasts and the cytosol ofS. tuberosum leaf cells. However, GS was specifically associated with the chloroplasts of mesophyll cells and with the cytoplasm of phloem companion cells. The evidence for cell-specific localization of chloroplast and cytosolic GS presented here agrees with the recently reported cell-specific pattern of expression of GUS reporter gene, directed by promoters for chloroplast and cytosolic GS form in tobacco transgenic plants. These data provide additional clues to the interpretation of the functional role of these different isoenzymes and its relationship with their specific localization.Abbreviations BSA
bovine serum albumin
- EM
electron microscope
- GOGAT
glutamate synthase
- GS
glutamine synthetase
- GUS
-glucuronidase
- IgG
immunoglobulin
- PBS
phosphate buffer saline
- SDS-PAGE
sodium dodecyl sulphate-polyacrylamide gel electrophoresis 相似文献
102.
Influenza virus M2 protein has ion channel activity. 总被引:68,自引:0,他引:68
The influenza virus M2 protein was expressed in Xenopus laevis oocytes and shown to have an associated ion channel activity selective for monovalent ions. The anti-influenza virus drug amantadine hydrochloride significantly attenuated the inward current induced by hyperpolarization of oocyte membranes. Mutations in the M2 membrane-spanning domain that confer viral resistance to amantadine produced currents that were resistant to the drug. Analysis of the currents of these altered M2 proteins suggests that the channel pore is formed by the transmembrane domain of the M2 protein. The wild-type M2 channel was found to be regulated by pH. The wild-type M2 ion channel activity is proposed to have a pivotal role in the biology of influenza virus infection. 相似文献
103.
Summary The production of itaconic acid by immobilizedAspergillus terreus TKK 200-5-1 was studied both in shake flask cultures, and in continuous column bioreactors. The effect of glucose and ammonium nitrate concentrations, and of pH were examined using a statistical experimental plan. The highest itaconic acid product concentration could be reached at the highest investigated glucose concentration of 150 g/l and the highest initial pH of 3.75, in the absence of ammonium nitrate. In a continuous packed bed column system operated fro 4.5 months itaconic acid was obtained at a productivity of 328 mg/d per gram of polyurethane foam carrier. 相似文献
104.
A case of Actinomyces infection of the breast secondary to thoracopleural disease was initially diagnosed by fine needle aspiration biopsy. The tender, hard, swollen left breast was clinically suspected of harboring an inflammatory carcinoma. Cell block sections of the aspirate showed polymorphonuclear leukocytes surrounding a typical "sulphur granule" composed of branching filaments. The diagnosis was confirmed by culture of material obtained at subsequent surgery. 相似文献
105.
Diagnosis of cystic pancreatic lesions by cytologic examination and carcinoembryonic antigen and amylase assays of cyst contents. 总被引:6,自引:0,他引:6
The contents obtained by fine needle aspiration (FNA) from 41 pancreatic cysts in 32 patients were studied cytologically and assayed for amylase and carcinoembryonic antigen (CEA) levels, which have been shown to discriminate pancreatic pseudocysts from mucinous cystic neoplasms and necrotic cystic carcinomas. The results were correlated with the histopathologic findings following surgery or with a clinical and radiologic follow-up of up to two years. The clinical, radiologic and cytologic characteristics did not discriminate pseudocysts from cystic neoplasms. The amylase content of cysts was high in pseudocysts, cystic carcinomas and mucinous cystic neoplasms. The mean CEA content was highest in cystic carcinomas and mucinous cysts and low in pseudocysts. The cytologic diagnosis of mucinous cystic neoplasms and carcinomas had a sensitivity of 54% and a specificity of 91%. The diagnosis of these lesions based on a CEA level greater than 10 ng/ml had a sensitivity of 100% and a specificity of 81%. The adjunctive use of CEA content analysis enhanced the sensitivity of the cytologic diagnosis of mucinous cystic neoplasms and carcinomas to 100%. 相似文献
106.
A fast purification procedure for the isolation and purification of eukaryotic porin (De Pinto et al., (1987) Biochim. Biophys. Acta 905, 499-502) was applied to liver mitochondria of the fish Anguilla anguilla. A protein preparation was obtained which formed slightly anionically selective pores in reconstitution experiments with lipid bilayer membranes. The distribution of single-channel conductances had two maxima of 2.4 nS and 4.0 nS in 1 M KCl. Sodium dodecylsulfate electrophoretograms of the protein preparation showed the presence of two bands of very similar electrophoretic mobility (32 and 32.5 kDa). Both bands cross-reacted with antibodies raised against purified bovine heart porin and with antibodies raised against the 19 amino acids N-terminal end of human porin. No cross-reactivity was observed with antibodies against yeast porin. The peptide maps of the two bands showed slight differences. The possibility of the presence of two different porins in liver mitochondria of Anguilla anguilla is discussed. An extensive immunological comparison of different mitochondrial porins is presented. 相似文献
107.
Purification and characterization of farnesyl pyrophosphate synthase from Capsicum annuum 总被引:5,自引:0,他引:5
Farnesyl pyrophosphate synthase (FPP) displaying dimethylallyl transferase activity (EC 2.5.1.1) and geranyl transferase activity (EC 2.5.1.10) was purified from Capsicum fruits. This prenyltransferase has a molecular mass of 89,000 +/- 5000 Da resulting from the association of two apparently identical subunits having a molecular mass of 43,000 +/- 2000 Da. Antibodies raised against Capsicum FPP synthase selectively blocked the transferase activity. Analysis of the immunological relationships between FPP synthase and geranylgeranyl pyrophosphate synthase (EC 2.5.1.1, EC 2.5.1.10 and EC 2.5.1.30) revealed that these two enzymes though performing the same mechanism of catalysis and accepting identical substrates have different antigenic determinants. Thus, in connection to previous work, this immunological study suggests that Capsicum FPP is strictly located in the extraplastidial compartment. 相似文献
108.
The role of positive charges located on the hydrophilic surface of the mitochondrial outer membrane channel was investigated by studying the interaction between LDAO-solubilized porin and a cation-exchanger column. The binding of porin to the column material was inhibited when the elution buffer had a pH of 9 or when 2 mM dextran sulfate was added to the buffer at neutral pH. Interestingly, the addition of a synthetic copolymer of methacrylate, maleate and styrene known as a potent modulator of the voltage-dependence, did not influence the interaction between column material and porin. Incubation of porin with fluorescein isothiocyanate (FITC) resulted in the isolation of a porin fraction in which on average two lysines located on the surface of the pore-forming complex per 35 kDa polypeptide were modified. The voltage-dependence of the fluorescein isothiocyanate modified porin was strongly decreased as compared with the unmodified porin. The experiments presented here give the first biochemical evidence that positively charged lysine residues located on the surface of the channel-forming complex are responsible for the gating of the mitochondrial porin-channel. 相似文献
109.
Graham V. Brown Ross L. Coppel Helena Vrbova Raelene J. Grumont Robin F. Anders 《Experimental parasitology》1982,53(2):279-284
Proteins of erythrocytic stages of Plasmodium falciparum were biosynthetically labeled at different times during the first cycle of in vitro synchronous cultivation after collection from patients in the Madang region of Papua New Guinea. Proteins were immunoprecipitated with a pool of hyperimmune serum collected in the region then analyzed by sodium dodecyl sulfate-gel electrophoresis. Antigens were recognized in all life cycle stages but the majority of antigens, particularly those of high molecular weight, were present in the mature forms of the parasite. 相似文献
110.