ABSTRACT
Bcl-2-associated X (BAX) protein plays a gatekeeper role in transmitting apoptotic signaling from cytosol to mitochondria. However, little is known about its stability. This study reports a comprehensive investigation on the stability of BAX using spin-label ESR, CD, and ThermoFluor methods. Point mutations covering all of the nine helices of BAX were prepared. ESR study shows that BAX can be divided into two structural regions, each responding differently to the presence of guanidine hydrochloride (GdnHCl). The N-terminal region (helices 1-3) is denatured in 6 M GdnHCl, whereas the C-terminal region (helices 4-9) is resistant to the denaturing effects. The far-UV CD spectra show an appreciable amount of helical content of BAX at high temperatures. The magnitude of the near-UV CD signal is increased with increasing temperature in either 0 or 6 M GdnHCl, indicating an enhancement of aromatic side-chain packing in the C-terminal region. Taken together with ThermoFluor results, we show that a core interior, wherein aromatic interactions are highly involved, within the C-terminal region plays an important role in stabilizing BAX against the denaturing effects. Collectively, we report a highly stable, indestructible intermediate state of BAX. Side-chain packing interactions are shown to be the major stabilizing force in determining BAX structure.
Subject(s)
Guanidine/chemistry , Temperature , bcl-2-Associated X Protein/chemistry , Humans , Models, Molecular , Protein DenaturationABSTRACT
In recent years, graphene studies have increased rapidly. Graphene oxide, which is an intermediate product to form graphene, is insulating, and it should be thermally reduced to be electrically conductive. We herein describe an attempt to make use of the insulating properties of graphene oxide. The graphene oxide layers are deposited onto Si substrates, and a metal-insulator-semiconductor tunneling structure is formed and its optoelectronic properties are studied. The accumulation dark current and inversion photocurrent of the graphene oxide device are superior to the control device. The introduction of graphene oxide improves the rectifying characteristic of the diode and enhances its responsivity as a photodetector. At 2 V, the photo-to-dark current ratio of the graphene oxide device is 24, larger than the value of 15 measured in the control device.
