Identification of HIV-1 nucleocapsid protein: nucleic acid antagonists with cellular anti-HIV activity

Stephen, Andrew; Worthy, Karen M.; Towler, Eric M.; Mikovits, Judy A.; Sei, Shizuko; Roberts, Paula; Yang, Quan-en; Akee, Rhone K.; Klausmeyer, Paul; McCloud, Thomas G.; Henderson, L.E.; Rein, Alan; Covell, David G.; Currens, Michael J.; Shoemaker, Robert H.; Fisher, Robert J.

doi:10.1016/s0006-291x(02)02063-6

Cited by 57 publications

(62 citation statements)

References 40 publications

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“…The concentration at which the activity of NCp7 was found to be negligible was used to obtain an approximate value for the effective dissociation constant of the compound to NCp7. For the rare chemical compounds received from DTP, the values estimated in this work are compared with the dissociation constants published by Stephen et al (27). With one exception, the values obtained in our experiments were mostly within one order of magnitude of the previously reported values.…”

Section: Resultssupporting

confidence: 76%

“…While it is possible that tetrachlorofluorescein is effective at concentrations higher than 10 μM (in 50 mM NaCl), such high concentrations would likely be toxic for the host cells, so they were not tested. In general, we were able to confirm that the fluorescein-like compounds are much less active than the gallein-like compounds, as reported in the previous study (27).…”

Section: Screening the Commercially-available Gallein-like And Fluoresupporting

confidence: 90%

“…Given its previously shown anti-retroviral activity (27), tetrachlorogallein behaved differently than expected in our assay. The effect on the DNA stretching curve was dependent not only on the compound concentration, but also on the incubation time.…”

Section: Screening the Commercially-available Gallein-like And Fluorementioning

confidence: 56%

“…However, the zinc ejector agents are still avoided because of their potential cytotoxicity at high concentrations. This study focuses on a small number of non-zinc-ejector compounds, identified in a previous study (27). These compounds were found by screening the influence of a large number of compounds on the efficiency of NC binding to d(TG) 4 oligos in a high throughput luminescence based assay and confirmed by BIAcore analysis.…”

Section: Introductionmentioning

confidence: 99%

“…These compounds were previously identified as reversible, non-covalent, non-redox binders, and were also shown to be non-zinc-ejectors. The experimental results refer to several compounds previously reported to have NCp7 antagonist activity (27), and to one example of a new series of compounds identified in recent studies by the Developmental Therapeutic Program (DTP), HIV Drug Resistance Program, National Cancer Institute (NCI), Frederick, MD, USA. It is worth noting that the new method reported here can be used to screen many types of compounds including fluorescent dyes, for which standard fluorescence titration assays can be strongly affected by a high fluorescence background.…”

Section: Introductionmentioning

confidence: 99%

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Single DNA molecule stretching measures the activity of chemicals that target the HIV-1 nucleocapsid protein

Cruceanu

Stephen

Beuning

et al. 2006

Analytical Biochemistry

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We develop a biophysical method for investigating chemical compounds that target the nucleic acid chaperone activity of HIV-1 nucleocapsid protein (NCp7). We used an optical tweezers instrument to stretch single λ-DNA molecules through the helix-to-coil transition in the presence of NCp7 and various chemical compounds. The change in the helix-coil transition width induced by wild-type NCp7 and its zinc finger variants correlates with in vitro nucleic acid chaperone activity measurements and in vivo assays. The compound-NC interaction measured here reduces NCp7's capability to alter the transition width. Purified compounds from the NCI Diversity set, 119889, 119911, and 119913 reduce the chaperone activity of 5 nM NC in aqueous solution at 10 nM, 25 nM, and 100 nM concentration, respectively. Similarly, gallein reduced the activity of 4 nM NC at 100 nM concentration. Further analysis allows us to dissect the impact of each compound on both sequence-specific and non-sequence-specific DNA binding of NC, two of the main components of NC's nucleic acid chaperone activity. These results suggest that DNA stretching experiments can be used to screen chemical compounds targeting NC proteins, and to further explore the mechanisms by which these compounds interact with NC and alter its nucleic acid chaperone activity.

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Section: Resultssupporting

confidence: 76%

Section: Screening the Commercially-available Gallein-like And Fluoresupporting

confidence: 90%

Section: Screening the Commercially-available Gallein-like And Fluorementioning

confidence: 56%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Single DNA molecule stretching measures the activity of chemicals that target the HIV-1 nucleocapsid protein

Cruceanu

Stephen

Beuning

et al. 2006

Analytical Biochemistry

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E

2010

Handbook of Biological Dyes and Stains

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O COONa O NaO O 2 N NO 2 Br BrCA Index Name Spiro[isobenzofuran-1(3H),9 0 -[9H] xanthen]-3-one, 4 0 ,5 0 -dibromo-3 0 ,6 0 -dihydroxy-2 0 ,7 0 -dinitro-, sodium salt (1 : 2) Other Names Fluorescein, 4 0 ,disodium salt; C.I. 45400; C.I. Acid Red 91; Dibromodinitrofluorescein sodium; Eosin B; Eosin BA; Eosine B; Eosine BA; Eosine BN; Eosine BNX; Eosine I Bluish; Saffrosine; Simacid Nitro Eosin 24669 Merck Index Number 3602 Chemical/Dye Class Xanthene Molecular Formula C 20 H 6 Br 2 N 2 Na 2 O 9 Molecular Weight 624.06 Physical Form Red-brown to green crystals or powder Solubility Freely soluble in water; soluble in ethanol Melting Point 295 C Absorption (l max ) 514 nm, 395 nm Emission (l max ) 544 nm Synthesis Synthetic method 1-4 Staining Applications Brain; 5 cells; 6 microorganisms; 7 nucleic acids; 8 peptides; 9 proteins; 9-13 enzyme substrates; 14,15 hairs 16 Sakamoto, H. Development of a colorimetric quantitation method of protein in solution by excluding the influence of creatinine. PCT Int. Appl. WO 2005071420, 2005; Chem. Abstr. 2005, 143, 188838. 11. Waheed, A. A.; Rao, K. S.; Gupta, P. D. Mechanism of dye binding in the protein assay using eosin dyes. Anal. Biochem. 2000, 287, 73-79. 12. Waheed, A. A.; Gupta, P. D. Application of an eosin B dye method for estimating a wide range of proteins. Wikborg, M. Pre-stained 3,3 0 ,5,5 0tetrame-thylbenzidine substrates for the detection of enzyme activity. PCT Int. Appl. WO 9904261, 1999; Chem. Abstr. 1999, 130, 150342. 15. Sumiya, M. Dyeing of substrates. Jpn. Kokai Tokkyo Koho JP 62021884, 1987; Chem. Abstr. 1987, 107, 117243. 16. Moeller, H.; Meinigke, B. Hair dye containing nitroxanthene derivatives. Eur. Pat. Appl. EP 1199065, 2002; Chem. Abstr. 2002, 136, 345464. 17. Massimine, K. M.; McIntosh, M. T.; Doan, L. T.; Atreya, C. E.; Gromer, S.; Sirawaraporn, W.; Elliott, D. A.; Joiner, K. A.; Schirmer, R. H.; Anderson, K. S. Eosin B as a novel antimalarial agent for drugresistant Plasmodium falciparum. Antimicrob. Agents Chemother. 2006, 50, 3132-3141. 18. Das Gupta, P.; Waheed, A. L. A.; A process for the preparation of a formulation useful for the detection and estimation of proteins present in biological fluids. Zimmermann, H.; Fritz-Wolf, K. Ellagic acid-related compound and polyaromatic phenol inhibitors of glutathione-S-transferase, and their therapeutic use. PCT Int.

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Endoplasmic Reticulum Targeting Reactive Oxygen Species Sensor Based on Dihydrofluorescein: Application of Endoplasmic Reticulum Stress

Lê

et al. 2020

Bulletin Korean Chem Soc

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Endoplasmic reticulum (ER) has a unique redox environment, which plays critical roles in the organelle's function and in its pathological responses such as ER stress. In this work, we introduce an ER-targeting fluorogenic reactive oxygen species (ROS) chemosensor (ER-Flu) from copper(I)-catalyzed alkyne-azide cycloaddition of 3-propargyl ester of 2 0 ,7 0-dichlorodihydrofluorescein diacetate and N 3-glibenclamide, which were adopted as a fluorogenic ROS sensing module and an ER-targeting module, respectively. Thereby, a series of confocal microscopic experiments of ER-Flu demonstrated that the sensor localizes in ER of the live cells and that ROS are elevated in the cells by ER stress inducers such as thapsigargin, brefeldin A, and tunicamycin.

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Identification of HIV-1 nucleocapsid protein: nucleic acid antagonists with cellular anti-HIV activity

Cited by 57 publications

References 40 publications

Single DNA molecule stretching measures the activity of chemicals that target the HIV-1 nucleocapsid protein

Single DNA molecule stretching measures the activity of chemicals that target the HIV-1 nucleocapsid protein

E

Endoplasmic Reticulum Targeting Reactive Oxygen Species Sensor Based on Dihydrofluorescein: Application of Endoplasmic Reticulum Stress

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