Membrane topology analysis of HIV-1 envelope glycoprotein gp41

<p>Abstract</p> <p>Background</p> <p>The gp41 subunit of the HIV-1 envelope glycoprotein (Env) has been widely regarded as a type I transmembrane protein with a single membrane-spanning domain (MSD). An alternative topology model suggested multiple MSDs. The major discrepancy between the two models is that the cytoplasmic Kennedy sequence in the single MSD model is assigned as the extracellular loop accessible to neutralizing antibodies in the other model. We examined the membrane topology of the gp41 subunit in both prokaryotic and mammalian systems. We attached topological markers to the C-termini of serially truncated gp41. In the prokaryotic system, we utilized a green fluorescent protein (GFP) that is only active in the cytoplasm. The tag protein (HaloTag) and a membrane-impermeable ligand specific to HaloTag was used in the mammalian system.</p> <p>Results</p> <p>In the absence of membrane fusion, both the prokaryotic and mammalian systems (293FT cells) supported the single MSD model. In the presence of membrane fusion in mammalian cells (293CD4 cells), the data obtained seem to support the multiple MSD model. However, the region predicted to be a potential MSD is the highly hydrophilic Kennedy sequence and is least likely to become a MSD based on several algorithms. Further analysis revealed the induction of membrane permeability during membrane fusion, allowing the membrane-impermeable ligand and antibodies to cross the membrane. Therefore, we cannot completely rule out the possible artifacts. Addition of membrane fusion inhibitors or alterations of the MSD sequence decreased the induction of membrane permeability.</p> <p>Conclusions</p> <p>It is likely that a single MSD model for HIV-1 gp41 holds true even in the presence of membrane fusion. The degree of the augmentation of membrane permeability we observed was dependent on the membrane fusion and sequence of the MSD.</p

The Acute Effects of a Single Dose of Molecular Hydrogen Supplements on Responses to Ergogenic Adjustments during High-Intensity Intermittent Exercise in Humans

This research examined the effects of single-dose molecular hydrogen (H2) supplements on acid-base status and local muscle deoxygenation during rest, high-intensity intermittent training (HIIT) performance, and recovery. Ten healthy, trained subjects in a randomized, double-blind, crossover design received H2-rich calcium powder (HCP) (1500 mg, containing 2.544 &mu;g of H2) or H2-depleted placebo (1500 mg) supplements 1 h pre-exercise. They performed six bouts of 7 s all-out pedaling (HIIT) at 7.5% of body weight separated by 40 s pedaling intervals, followed by a recovery period. Blood gases&rsquo; pH, PCO2, and HCO3&minus; concentrations were measured at rest. Muscle deoxygenation (deoxy[Hb + Mb]) and tissue O2 saturation (StO2) were determined via time-resolved near-infrared spectroscopy in the vastus lateralis (VL) and rectus femoris (RF) muscles from rest to recovery. At rest, the HCP group had significantly higher PCO2 and HCO3&minus; concentrations and a slight tendency toward acidosis. During exercise, the first HIIT bout&rsquo;s peak power was significantly higher in HCP (839&thinsp; &plusmn; &thinsp;112 W) vs. Placebo (816&thinsp; &plusmn; &thinsp;108 W, p = 0.001), and HCP had a notable effect on significantly increased deoxy[Hb + Mb] concentration during HIIT exercise, despite no differences in heart rate response. The HCP group showed significantly greater O2 extraction in VL and microvascular (Hb) volume in RF during HIIT exercise. The HIIT exercise provided significantly improved blood flow and muscle reoxygenation rates in both the RF and VL during passive recovery compared to rest in all groups. The HCP supplement might exert ergogenic effects on high-intensity exercise and prove advantageous for improving anaerobic HIIT exercise performance

Effects of High-Intensity Anaerobic Exercise on the Scavenging Activity of Various Reactive Oxygen Species and Free Radicals in Athletes

High-intensity exercise in athletes results in mainly the production of excess reactive oxygen species (ROS) in skeletal muscle, and thus athletes should maintain greater ROS scavenging activity in the body. We investigated the changes in six different ROS-scavenging activities in athletes following high-intensity anaerobic exercise. A 30-s Wingate exercise test as a form of high-intensity anaerobic exercise was completed by 10 male university track and field team members. Blood samples were collected before and after the exercise, and the ROS-scavenging activities (OH•, O2•−, 1O2, RO• and ROO•, and CH3•) were evaluated by the electron spin resonance (ESR) spin-trapping method. The anaerobic exercise significantly increased RO• and ROO• scavenging activities, and the total area of the radar chart in the ROS-scavenging activities increased 178% from that in pre-exercise. A significant correlation between the mean power of the anaerobic exercise and the 1O2 scavenging activity was revealed (r = 0.72, p n = 5, each). These results suggest that (i) the scavenging activities of some ROS are increased immediately after high-intensity anaerobic exercise, and (ii) an individual’s OH• scavenging activity responsiveness may be related to his anaerobic exercise performance. In addition, greater pre-exercise 1O2 scavenging activity might lead to the generation of higher mean power in high-intensity anaerobic exercise