Interflap Distances in HIV-1 Protease Determined by Pulsed EPR Measurements

2007 ◽  
Vol 129 (36) ◽  
pp. 11004-11005 ◽  
Author(s):  
Luis Galiano ◽  
Marco Bonora ◽  
Gail E. Fanucci
Keyword(s):  
Pulsed Epr ◽  
Epr Measurements ◽  
Hiv 1

scholarly journals Pulsed EPR characterization of HIV-1 protease conformational sampling and inhibitor-induced population shifts

2016 ◽  
Vol 18 (8) ◽  
pp. 5819-5831 ◽  
Author(s):  
Zhanglong Liu ◽  
Thomas M. Casey ◽  
Mandy E. Blackburn ◽  
Xi Huang ◽  
Linh Pham ◽  
...  
Keyword(s):  
Spin Labeling ◽  
Conformational Sampling ◽  
Electron Resonance ◽  
Pulsed Epr ◽  
Double Electron ◽  
Conformational Landscape ◽  
Double Electron Electron Resonance ◽  
Hiv 1

The conformational landscape of HIV-1 protease can be characterized by double electron–electron resonance (DEER) spin-labeling.

Conformational landscape of non-B variants of HIV-1 protease: A pulsed EPR study

2020 ◽  
Vol 532 (2) ◽  
pp. 219-224 ◽  
Author(s):  
Trang T. Tran ◽  
Zhanglong Liu ◽  
Gail E. Fanucci
Keyword(s):  
Pulsed Epr ◽  
Conformational Landscape ◽  
Hiv 1

scholarly journals Inhibitor-Induced Conformational Shifts and Ligand-Exchange Dynamics for HIV-1 Protease Measured by Pulsed EPR and NMR Spectroscopy

2012 ◽  
Vol 116 (49) ◽  
pp. 14235-14244 ◽  
Author(s):  
Xi Huang ◽  
Ian Mitchelle S. de Vera ◽  
Angelo M. Veloro ◽  
Mandy E. Blackburn ◽  
Jamie L. Kear ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Ligand Exchange ◽  
Pulsed Epr ◽  
Hiv 1 ◽  
Conformational Shifts

Monitoring Inhibitor-Induced Conformational Population Shifts in HIV-1 Protease by Pulsed EPR Spectroscopy

Biochemistry ◽  
2009 ◽  
Vol 48 (37) ◽  
pp. 8765-8767 ◽  
Author(s):  
Mandy E. Blackburn ◽  
Angelo M. Veloro ◽  
Gail E. Fanucci
Keyword(s):  
Epr Spectroscopy ◽  
Pulsed Epr ◽  
Conformational Population ◽  
Hiv 1

scholarly journals Probing the Y2 Receptor on Transmembrane, Intra- and Extra-Cellular Sites for EPR Measurements

Molecules ◽  
2020 ◽  
Vol 25 (18) ◽  
pp. 4143
Author(s):  
Jeannette M. Laugwitz ◽  
Haleh H. Haeri ◽  
Anette Kaiser ◽  
Ulrike Krug ◽  
Dariush Hinderberger ◽  
...  
Keyword(s):  
Continuous Wave ◽  
Conformational Dynamics ◽  
Spin Labeling ◽  
Conformational States ◽  
Distance Measurements ◽  
Paramagnetic Resonance ◽  
Pulsed Epr ◽  
Y2 Receptor ◽  
Epr Measurements ◽  
Double Electron Electron Resonance

The function of G protein-coupled receptors is intrinsically linked to their conformational dynamics. In conjugation with site-directed spin labeling, electron paramagnetic resonance (EPR) spectroscopy provides powerful tools to study the highly dynamic conformational states of these proteins. Here, we explored positions for nitroxide spin labeling coupled to single cysteines, introduced at transmembrane, intra- and extra-cellular sites of the human neuropeptide Y2 receptor. Receptor mutants were functionally analyzed in cell culture system, expressed in Escherichia coli fermentation with yields of up to 10 mg of purified protein per liter expression medium and functionally reconstituted into a lipid bicelle environment. Successful spin labeling was confirmed by a fluorescence assay and continuous wave EPR measurements. EPR spectra revealed mobile and immobile populations, indicating multiple dynamic conformational states of the receptor. We found that the singly mutated positions by MTSL ((1-oxyl-2,2,5,5-tetramethyl-2,5-dihydro-1H-pyrrol-3-yl) methyl methanesulfonothioate) have a water exposed immobilized conformation as their main conformation, while in case of the IDSL (bis(1-oxyl-2,2,5,5-tetramethyl-3-imidazolin-4-yl) disulfide) labeled positions, the main conformation are mainly of hydrophobic nature. Further, double cysteine mutants were generated and examined for potential applications of distance measurements by double electron–electron resonance (DEER) pulsed EPR technique on the receptor.

scholarly journals Dielectric Resonator for K a-Band Pulsed EPR Measurements at Cryogenic Temperatures: Probehead Construction and Applications

2012 ◽  
Vol 42 (4) ◽  
pp. 441-452 ◽  
Author(s):  
A. Raitsimring ◽  
A. Astashkin ◽  
J. H. Enemark ◽  
A. Blank ◽  
Y. Twig ◽  
...  
Keyword(s):  
Dielectric Resonator ◽  
Cryogenic Temperatures ◽  
Pulsed Epr ◽  
Epr Measurements

Ultrastructural observations of human monocytes infected with HIV-1

1991 ◽  
Vol 49 ◽  
pp. 108-109
Author(s):  
James K. Koehler ◽  
Steven G. Reed ◽  
Joao S. Silva
Keyword(s):  
Gradient Centrifugation ◽  
Virus Production ◽  
Human Monocyte ◽  
Red Cross ◽  
Human Monocytes ◽  
Blood Monocytes ◽  
Hiv Replication ◽  
Electron Microscopic ◽  
Ultrastructural Studies ◽  
Hiv 1

As part of a larger study involving the co-infection of human monocyte cultures with HIV and protozoan parasites, electron microscopic observations were made on the course of HIV replication and infection in these cells. Although several ultrastructural studies of the cytopathology associated with HIV infection have appeared, few studies have shown the details of virus production in “normal,” human monocytes/macrophages, one of the natural targets of the virus, and suspected of being a locus of quiescent virus during its long latent period. In this report, we detail some of the interactions of developing virons with the membranes and organelles of the monocyte host.Peripheral blood monocytes were prepared from buffy coats (Portland Red Cross) by Percoll gradient centrifugation, followed by adherence to cover slips. 90-95% pure monocytes were cultured in RPMI with 5% non-activated human AB serum for four days and infected with 100 TCID50/ml of HIV-1 for four hours, washed and incubated in fresh medium for 14 days.

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