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11.
New methods to investigate ATP requirement for pre-mRNA splicing: inhibition by hexokinase/glucose or an ATP-binding site blocker 总被引:1,自引:0,他引:1
Addition of yeast hexokinase and glucose at various time points of the pre-mRNA splicing reaction rapidly depleted ATP and inhibited further progress of the reaction, indicating that ATP is required for both the first and second steps of splicing. ATP analogues, p-fluorosulfonylbenzoyl-5'-adenosine (FSBA) and 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl), which can modify amino acids at the ATP-binding site of a protein, inactivated the splicing activity of the nuclear extract. While the inactivation by the former was irreversible, the splicing activity was complemented by a Micrococcal nuclease-treated extract. This ATP analogue (FSBA) may be a useful tool for identification of ATP-dependent splicing factors. 相似文献
12.
13.
Pure phloem sap was collected from leaf sheaths of Zea maysL. by the insect laser technique, and its chemical compositionwas analyzed. Sucrose was the only sugar detected. The predominantinorganic ions were K+ and Cl. The adenylate energy chargeof phloem sap was between 0.72 and 0.86. (Received October 18, 1989; Accepted May 11, 1990) 相似文献
14.
H Ohshima S Oguchi H Adachi S Iida H Suzuki T Sugimura H Esumi 《Biochemical and biophysical research communications》1992,183(1):238-244
Nitric oxide (NO) synthase (EC 1.14.23) was purified to homogeneity from bovine cerebrum. The molecular weight of NO synthase was estimated to be 150 kDa by both SDS/PAGE and gel filtration at high salt concentration. For activity, the enzyme required NADPH, Ca2+, calmodulin and tetrahydrobiopterin as cofactors. Rabbit polyclonal antibody to bovine brain NO synthase reacted with 150 kDa NO synthase in various bovine and rat organs, including the brain, pituitary and adrenal glands, but not with that in stimulated macrophages, indicating that there are at least two immunologically distinct NO synthases. 相似文献
15.
Application of the micropipette technique to the measurement of cultured porcine aortic endothelial cell viscoelastic properties 总被引:13,自引:0,他引:13
M Sato D P Theret L T Wheeler N Ohshima R M Nerem 《Journal of biomechanical engineering》1990,112(3):263-268
The viscoelastic deformation of porcine aortic endothelial cells grown under static culture conditions was measured using the micropipette technique. Experiments were conducted both for control cells (mechanically or trypsin detached from the substrate) and for cells in which cytoskeletal elements were disrupted by cytochalasin B or colchicine. The time course of the aspirated length into the pipette was measured after applying a stepwise increase in aspiration pressure. To analyze the data, a standard linear viscoelastic half-space model of the endothelial cell was used. The aspirated length was expressed as an exponential function of time. The actin microfilaments were found to be the major cytoskeletal component determining the viscoelastic response of endothelial cells grown in static culture. 相似文献
16.
Hiroyuki Tsunoda Toshio Ohshima Jun Tohyama Masayuki Sasaki Norio Sakuragawa Frank Martiniuk 《Human genetics》1996,97(4):496-499
We report a missense mutation in an adult Japanese patient with acid alpha-glucosidase (GAA) deficiency. A TC to GT transition
at nucleotides 1585–1586, was identified. This transition resulted in an amino acid substitution of Ser-529 to Val (S529V)
in exon 11. We also have demonstrated that the S529V mutation abolishes the catalytic activity of the enzyme. Our data suggest
that this mutation is the cause of the clinical manifestation known as adult-onset GAA deficiency. The missense mutation described
here is a new mutation, and the first identified in Japanese patients with GAA deficiency.
Received: 23 May 1995 相似文献
17.
Mitsuo Matsuda Toshiya Nosaka Masaaki Sato Norio Ohshima 《European journal of applied physiology and occupational physiology》1993,66(2):122-126
To evaluate the effects of exercise on aortic wall elasticity and elastic components, young male rats underwent various exercise regimes for 16 weeks. In the exercised rats, the aortic incremental elastic modulus decreased significantly when under physiological strain. The aortic content of elastin increased significantly and the calcium content of elastin decreased significantly in the exercised group. The accumulated data from the exercised and sedentary groups revealed that the elastin calcium content was related positively to the incremental elastic modulus. We concluded that physical exercise from an early age decreases the calcium deposit in aortic wall elastin and that this effect probably produced in the exercised rats a distensible aorta. 相似文献
18.
Koichi Ohshima Masahiro Kikuchi Shinichi Kobari Yuichi Masuda Fuyuki Eguchi Nobuhiro Kimura 《Virchows Archiv. B, Cell pathology including molecular pathology》1993,63(1):197-198
Using the polymerase chain reaction (PCR) to examine the occurrence ofbcl-2/JH joining produced by t(14;18) chromosomal translocation, amplified DNA was detected in 2 of 18 lymph nodes showing reactive
lymphadenopathy. The PCR was repeated in these two lymphs nodes using the same DNA samples, but no amplification was detected
at the second attempt. Thus the amplified DNA was considered to be derived from one copy of joinedbcl-2/JH in one cell, or from a few copies in a few clonal cells with the same joinedbcl-2/JH. These results suggest that false joining ofbcl-2/JH at the t(14;l8) junction may occur in reactive lymph nodes. 相似文献
19.
Histochemical assessment of selected carbohydrate sequences on Langerhans cells of human oral mucosa was made by combined use of enzyme digestion and immunostain-ing with monoclonal antibodies against specific carbohydrate structures. In both frozen sections and epithelial sheets without the enzyme pretreatment, mucosal Langerhans cells, identified by positive staining with anti-CD1a and HLA-DR antibodies, did not express any carbohydrate antigens on their surface. In contrast, following neuraminidase pretreatment of both types of material, the fucosylated type 2 chain (LeX) became detectable on Langerhans cells, indicating that sialic acid is the terminal residue of this sequence. Other enzymes were ineffective in this apparent unmasking, and the staining patterns of the other related carbohydrate sequences (Ley. Lea, Leb) remained unaffected by pretreatment with any of the enzymes used. These findings suggest that the mucosal Langerhans cells possess a unique carbohydrate chain, the sialyl fucosylated type 2 sequence (sialyl LeX antigen). 相似文献
20.