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91.
Stanislav D. Zakharov Xia Li Taya P. Red'ko Richard A. Dilley 《Journal of bioenergetics and biomembranes》1996,28(6):483-494
The 8-kDa subunit c of theE. coli F0 ATP-synthase proton channel was tested for Ca++ binding activity using a45Ca++ ligand blot assay after transferring the protein from SDS-PAGE gels onto polyvinyl difluoride membranes. The purified subunit c binds45Ca++ strongly with Ca++ binding properties very similar to those of the 8-kDa CF0 subunit III of choloroplast thylakoid membranes. The N-terminal f-Met carbonyl group seems necessary for Ca++ binding capacity, shown by loss of Ca++ binding following removal of the formyl group by mild acid treatment. The dicyclohexylcarbodiimide-reactive Asp-61 is not involved in the Ca++ binding, shown by Ca++ binding being retained in twoE. coli mutants, Asp61Asn and Asp61Gly. The Ca++ binding is pH dependent in both theE. coli and thylakoid 8-kDa proteins, being absent at pH 5.0 and rising to a maximum near pH 9.0. A treatment predicted to increase the Ca++ binding affinity to its F0 binding site (chlorpromazine photoaffinity attachment) caused an inhibition of ATP formation driven by a base-to-acid pH jump in whole cells. Inhibition was not observed when the Ca++ chelator EGTA was present with the cells during the chlorpromazine photoaffinity treatment. An apparent Ca++ binding constant on the site responsible for the UV plus chlorpromazine effect of near 80–100 nM was obtained using an EGTA-Ca++ buffer system to control free Ca++ concentration during the UV plus chlorpromazine treatment. The data are consistent with the notion that Ca++ bound to the periplasimic side of theE. coli F0 proton channel can block H+ entry into the channel. A similar effect occurs in thylakoid membranes, but the Ca++ binding site is on the lumen side of the thylakoid, where Ca++ binding can modulate acid-base jump ATP formation. The Ca++ binding to the F0 and CF0 complexes is consistent with a pH-dependent gating mechanism for control of H+ ion flux across the opening of the H+ channel.This work was supported in part by grants from the Department of Energy and the U.S. Department of Agriculture.On leave from the Institute of Soil Science and Photosynthesis, Russian Academy of Science, Pushchino, Russia. 相似文献
92.
In the search for candidate genes for the tuberous sclerosis (TSC1) disease locus on chromosome 9q34, we have isolated an overlapping series of 22 plasmid and phage cDNA clones covering nearly 7 kb and with an open reading frame of 5070 bp encoding a protein of 1690 amino acids. The putative protein product is a member of the kinesin superfamily and is homologous to the mouse KIF1A and theCaenorhabditas elegansunc-104 genes. Both KIF1A and unc-104 function in the anterograde axonal transport of synaptic vesicles. The human homolog is therefore termed H-ATSV (axonal transporter of synaptic vesicles, HGMW-approved nomenclature ATSV) Screening of DNA from 107 tuberous sclerosis patients and 80 unaffected individuals with H-ATSV cDNA probes by pulsed-field gel electrophoresis/Southern blotting following digestion by rare-cutting methylation-sensitive restriction enzymes showed variant banding patterns in three patients with tuberous sclerosis. However, further analysis indicated that these variant fragments represent a rare polymorphism probably associated with methylation of clustered restriction sites. There is no evidence to support H-ATSV as a candidate gene for TSC1. 相似文献
93.
Activation of recombinant trp by thapsigargin in Sf9 insect cells 总被引:12,自引:0,他引:12
94.
A systematic approach is presented to obtain the best door gap quality through optimal door fitting in automobile body manufacturing. First, three indexes of gap quality are defined; they are: (1) door gap width deviation relative to design nominal; (2) door gap parallelism; and (3) car-to-car gap consistency. Then the door-fitting problem is formulated into a general constrained optimization problem. The effects of optimal door fitting on the three quality indexes are evaluated through computer simulation. These results provide a lower bound on the design of nominal door gap by considering process capability. Finally, a computer-aided fixture adjustment scheme is developed to orient a door in a body side opening to achieve the optimal fitting. The amount of adjustment, with the desired orientation obtained from optimization, is calculated based on parametrically modeled local surface features of the fixture and the door. The adequacy of door feature modeling is verified through a door-fitting experiment. 相似文献
95.
96.
零下低温对杂交杨树皮层膜脂组成的影响 总被引:3,自引:0,他引:3
以不耐寒的美洲黑杨(Populusdeltoidescv.“Lux”I-69/55,父本)和耐寒性较强的欧美杨(P.euramericanaclcv.I-45/51,母本)的4个杂交F_1代无性系(95杨、559杨、600杨和1381杨)为材料,分析了零下低温寒潮前后枝条皮层的脂质组成。结果表明,寒潮影响下,皮层中磷脂含量增加而组成基本不变,膜脂脂肪酸组成的变化规律是:寒潮前脂肪酸不饱和指数(IUFA)值大的无性系,寒潮前后的IUFA值变化量小;寒潮前IUFA值较小的无性系,寒潮前后IUFA值变化量较大。本文借用力学概念,提出相对抗性概念,给出杨树无性系的相对抗性序列。序列表明F_1代无性系的耐寒性已较不耐寒的父本提高,这与田间观察基本一致。 相似文献
97.
莱氏衣原体膜上Mg~(2+)-ATPase用DOC溶解后,经Sepharose-6B和DEAE-CelluloseDE-52离子交换柱,得到了部分纯化的Mg~(2+)ATPase,并将此ATPase与不同极性头部的磷脂和膜糖脂重组,研究了不同的极性头部的磷脂和膜糖脂对ATPase活性的影响。此酶的活性不依赖酸性磷脂,PG、DPG、大豆磷脂等明显抑制酶活性,中性磷脂DMPC、PE、PC则能增加酶活性,其中尤以非双层脂PE的作用最为明显。从莱氏衣原体膜上提取的糖脂(MGDG,DGDG)单独和ATPase重组时,酶活性增加并不明显,当MGDG和DGDG以等比例混合时,能大大地增加酶活性。这表明Mg~(2+)-ATPase的活性很大程度上与磷脂的表面电荷及磷脂的组成相关。 相似文献
98.
Recombinant human kidney epithelial 293 cells were cultivated as aggregates in suspension. The concentration calcium ion, in the range of 100 muM to 1mM, affected the rate of aggregate formation. During the course of cultivation the size distribution of aggregates shifted and the fraction of larger aggregates increased. This effect was more profound in cultures with a high calcium concentration. Scanning and transmission microscopic examination of the aggregates revealed that cell packing was greater in the high calcium cultures and that ultrastructural integrity was retained in aggregates from both low and high calcium cultures. Confocal microscopy was applied to examine the viability of cells in the interior of the aggregates. High viability was observed in the aggregates obtained from exponentially growing cultures. Aggregates from the high calcium culture in the stationary phase exhibited lower viability in the interior. With its ease of retention in a perfusion bioreactor, aggregate cultures offer an alternative choice for large-scale operation. (c) 1993 John Wiley & Sons, Inc. 相似文献
99.
Cheng Jianxin Xia Yuqing Zhou Cheng Li Xiaohao Liu Pengfei 《Marine biotechnology (New York, N.Y.)》2023,25(2):291-313
Marine Biotechnology - Takifugu rubripes is important commercially fish species in China and it is under serious threat from white spot disease (cyptocaryoniasis), which leads to heavy economic... 相似文献
100.
Wang Zunxin Wang Xianyu Liu Siqin Yang Ying Li Yang Chen Siyuan Wang Guangpeng Zhang Xincheng Ye Yuxiu Hu Laibao Zhou Qing Wang Feibing Chen Xinhong 《Journal of Plant Growth Regulation》2023,42(1):294-303
Journal of Plant Growth Regulation - Zinc is an important micronutrient for the growth and development of human body and plants. Proper use of nitrogen fertilizer and foliar application of Zn have... 相似文献