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Title: F F{sub 1}-ATPase as biosensor to detect single virus

Abstract

F F{sub 1}-ATPase within chromatophore was constructed as a biosensor (immuno-rotary biosensor) for the purpose of capturing single virus. Capture of virus was based on antibody-antigen reaction. The detection of virus based on proton flux change driven by ATP-synthesis of F F{sub 1}-ATPase, which was indicated by F1300, was directly observed by a fluorescence microscope. The results demonstrate that the biosensor loading of virus particles has remarkable signal-to-noise ratio (3.8:1) compared to its control at single molecular level, and will be convenient, quick, and even super-sensitive for detecting virus particles.

Authors:
 [1];  [2];  [1];  [3];  [1];  [4]
  1. The National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China)
  2. (China)
  3. The National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101 (China). E-mail: yuejc@sun5.ibp.ac.cn
  4. The Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049 (China)
Publication Date:
OSTI Identifier:
20798910
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 342; Journal Issue: 4; Other Information: DOI: 10.1016/j.bbrc.2006.02.103; PII: S0006-291X(06)00357-3; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIBODIES; ANTIGENS; ATP; ATP-ASE; BIOSYNTHESIS; FLUORESCENCE; INFLUENZA VIRUSES; MICROSCOPES; SIGNAL-TO-NOISE RATIO

Citation Formats

Liu, XiaoLong, The Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, Zhang, Yun, Yue, JiaChang, Jiang, PeiDong, and Zhang, ZhenXi. F F{sub 1}-ATPase as biosensor to detect single virus. United States: N. p., 2006. Web. doi:10.1016/j.bbrc.2006.02.103.
Liu, XiaoLong, The Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, Zhang, Yun, Yue, JiaChang, Jiang, PeiDong, & Zhang, ZhenXi. F F{sub 1}-ATPase as biosensor to detect single virus. United States. doi:10.1016/j.bbrc.2006.02.103.
Liu, XiaoLong, The Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, Zhang, Yun, Yue, JiaChang, Jiang, PeiDong, and Zhang, ZhenXi. Fri . "F F{sub 1}-ATPase as biosensor to detect single virus". United States. doi:10.1016/j.bbrc.2006.02.103.
@article{osti_20798910,
title = {F F{sub 1}-ATPase as biosensor to detect single virus},
author = {Liu, XiaoLong and The Key Laboratory of Biomedical Information Engineering of Education Ministry, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049 and Zhang, Yun and Yue, JiaChang and Jiang, PeiDong and Zhang, ZhenXi},
abstractNote = {F F{sub 1}-ATPase within chromatophore was constructed as a biosensor (immuno-rotary biosensor) for the purpose of capturing single virus. Capture of virus was based on antibody-antigen reaction. The detection of virus based on proton flux change driven by ATP-synthesis of F F{sub 1}-ATPase, which was indicated by F1300, was directly observed by a fluorescence microscope. The results demonstrate that the biosensor loading of virus particles has remarkable signal-to-noise ratio (3.8:1) compared to its control at single molecular level, and will be convenient, quick, and even super-sensitive for detecting virus particles.},
doi = {10.1016/j.bbrc.2006.02.103},
journal = {Biochemical and Biophysical Research Communications},
number = 4,
volume = 342,
place = {United States},
year = {Fri Apr 21 00:00:00 EDT 2006},
month = {Fri Apr 21 00:00:00 EDT 2006}
}
  • In order to observe mechanically driven proton flux in F F{sub 1}-ATPase coupled with artificial driven rotation on F{sub 1} simultaneously, a double channel observation system was established. An artificial {delta}-free F F{sub 1}-ATPase was constructed with {alpha}{sub 3}, {beta}{sub 3}, {epsilon}, {gamma}, and c {sub n} subunits as rotator and a, b{sub 2} as stator. The chromatophore was immobilized on the glass surface through biotin-streptavidin-biotin system, and the magnetic bead was attached to the {beta} subunit of {delta}-free F F{sub 1}-ATPase. The mechanically driven proton flux was indicated by the fluorescence intensity change of fluorescein reference standard (F1300) andmore » recorded by a cooled digital CCD camera. The mechanochemical coupling stoichiometry between F and F{sub 1} is about 4.15 {+-} 0.2H{sup +}/rev when the magnetic field rotated at 0.33 Hz (rps)« less
  • A Nanodevice was constructed by {delta}-free F{sub o}F{sub 1}-ATPase within chromatophores and actin filaments through biotinlipid-streptavidin-biotin-(AC{sub 5}){sub 2}Sulfo-OSu system. One actin filament linking with many chromatophores functions as the Nanodevice body and many {delta}-free F{sub o}F{sub 1}-ATPase as the Nanodevice motors. Movement of the Nanodevice was observed directly by fluorescence microscopy with CCD camera after illumination. The moving speed was about 2.17-24.43 {mu}m/s for various length Nanodevices and most of them were stopped by adding CCCP. This means that the Nanodevice was driven by PMF (proton-motive force) in the cooperating {delta}-free F{sub o}F{sub 1}-ATPase. From bioengineering point of view, themore » cooperation of F{sub o}F{sub 1}-ATPase is a very important research field in the future.« less
  • F{sub o}F{sub 1}-ATPase activity is regulated by external links on {beta} subunits with different molecular weight. It is inhibited when anti-{beta} subunit antibody, streptavidin and H9 antibody link on the {beta} subunits successively, but is activated when virus was binded. Western blotting indicated that the employed anti-{beta} antibody target was on the non-catalytic site of the {beta} subunit. Furthermore, an ESR study of spin-labeled ATP (SL-ATP) showed that the affinity of ATP to the holoenzyme increases with increasing external links on the {beta} subunits. This simple regulation method may have great potential in the design of rapid, free labeled, sensitivemore » and selective biosensors.« less
  • The recent finding that the presence of ATP at noncatalytic sites of chloroplast F[sub 1]-ATPase (CF[sub 1]) is necessary for ATPase activity prompted more detailed studies of the effect of noncatalytic site nucleotides on catalysis. CF[sub 1] containing at noncatalytic sites less than one ADP or about two ATP was prepared by heat activation in the absence of Mg[sup 2+] and in the presence of ADP or ATP, respectively. After removal of medium nucleotides, the CF[sub 1] preparations were used for measurement of the time course of nucleotide binding from 10 to 100 [mu]M concentrations of [sup 3]H-labeled ADP, ATP,more » or GTP. The presence of Mg[sup 2+] strongly promotes the tight binding of ADP and ATP at noncatalytic sites. For example, the ADP and ATP at noncatalytic sites. For example, the ADP-heat-activated enzyme in presence of 1 mM Mg[sup 2+] binds ADP with a rate constant of 0.5 [times] 10[sup 6] M[sup [minus]1] min[sup [minus]1] to give an enzyme with two ADP at noncatalytic sites with a K[sub d] of about 0.1 [mu]M. Upon exposure to Mg[sup 2+] and ATP the vacant noncatalytic site binds an ATP rapidly and, as an ADP slowly dissociates, a second ATP binds. The binding correlates with an increase in the ATPase activity. In contrast the tight binding of [[sup 3]H]GTP to noncatalytic sites gives an enzyme with no ATPase activity. The three noncatalytic sites differ in their binding properties. The noncatalytic site that remains vacant after the ADP-heat-activated CF[sub 1] is exposed to Mg[sup 2+] and ADP and that can bind ATP rapidly as designated as site A; the site that fills with ATP as ADP dissociates when then enzyme is exposed to Mg[sup 2+] and ATP is called site B, and the site to which ADP remains bound is called site C. Procedures are given for attaining CF[sub 1] with ADp at sites B and C, with GTP at sites A and/or B, and with ATP at sites A, B, and/or C, and catalytic activities of such preparations are measured.« less
  • 2-Azidoadenine (/sup 32/P)nucleotide was bound specifically at catalytic or noncatalytic nucleotide binding sites on beef heart mitochondrial F/sub 1/ ATPase. In both cases, photolysis resulted in nearly exclusive labeling of the ..beta.. subunit. The modified enzyme was digested with trypsin, and labeled peptides were purified by reversed-phase high-pressure liquid chromatography. Amino acid sequence analysis of the major /sup 32/P-labeled tryptic fragments showed ..beta..-subunit Tyr-368 to be present at noncatalytic sites and ..beta.. Tyr-345 to be present at catalytic sites. From the relationship between the degree of inhibition and extent of modification, it is estimated that one-third of the catalytic sitesmore » or two-thirds of the noncatalytic sites must be modified to give near-complete inhibition of catalytic activity.« less