New tetrameric forms of the rotavirus NSP4 with antiparallel helices

Rotavirus nonstructural protein 4, the first viral enterotoxin to be identified, is a multidomain, multifunctional glycoprotein. Earlier, we reported a Ca2+-bound coiled-coil tetrameric structure of the diarrhea-inducing region of NSP4 from the rotavirus strains SA11 and I321 and a Ca2+-free pentameric structure from the rotavirus strain ST3, all with a parallel arrangement of alpha-helices. pH was found to determine the oligomeric state: a basic pH favoured a tetramer, whereas an acidic pH favoured a pentamer. Here, we report two novel forms of the coiled-coil region of NSP4 from the bovine rotavirus strains MF66 and NCDV. These crystallized at acidic pH, forming antiparallel coiled-coil tetrameric structures without any bound Ca2+ ion. Structural and mutational studies of the coiled-coil regions of NSP4 revealed that the nature of the residue at position 131 (Tyr/His) plays an important role in the observed structural diversity

An in-vitro Cytotoxic and Genotoxic Properties of Allmanda Cathartica L. Latex Green NPs on Human Peripheral Blood Mononuclear Cells

Green synthesis of silver nanoparticles (NPs) by green route approaches has advantages over conventional methods. In green synthesis, we use eco-friendly plant extracts contain secondary metabolites and bioactive components, proteins that act as both reducing and capping agents, form stable and shape-controlled green silver nanoparticles. The current study deals with the synthesis of silver nanoparticles using the aqueous latex extract of Allmanda cathartica. The green silver nanoparticles are characterized by using different spectroscopic methods like ultra violet-visible spectroscopy (UV-Vis), Fourier transform-infrared spectroscopy (FTIR), transmission electron microscope (TEM), scanning electron microscope (SEM) and X-ray diffraction (XRD). Results indicated that the crystalline natured particles were spherical shaped with an average of 35 nm in size, and that the stability of silver nanoparticles was due to its high negative zeta potential of –27.6 mV. The current study also revealed that green silver nanoparticles had very good genotoxic and cytotoxic activity in peripheral blood mononuclear cells (PBMCs). Leukemia leads to the development of high numbers of white blood cells, which is one of the major types of cancers that affect children. Many of the chemicals used for the treatment produce remarkable side effects. To overcome this problem, we made an attempt to see the efficacy of latex green silver nanoparticle on peripheral blood mononuclear cells and deoxyribonucleic acid fragmentation, which leads to the development of future therapeutic drugs

The flexible C terminus of the rotavirus non-structural protein NSP4 is an important determinant of its biological properties

The rotavirus non-structural protein NSP4 functions as the viral enterotoxin and intracellular receptor for the double-layered particles (DLP). The full-length protein cannot be expressed and/or purified to homogeneity from bacterial or insect cells. However, a bacterially expressed and purified mutant lacking the N-terminal 72 aa(\triangle N72) was recently obtained from strains Hg18 and SA11 exhibiting approximately 17–20-, 150–200- and 13166–15800-fold lower $DD_{50}$ (50% diarrhoea-inducing dose) values in suckling mice compared with that reported for the partially pure, full-length protein, a C-terminal M175I mutant and a synthetic peptide comprising aa 114– 135, respectively, suggesting the requirement for a unique conformation for optimal functions of the purified protein. The stretch of approximately 40 aa from the C terminus of the cytoplasmic tail of the endoplasmic reticulum-anchored NSP4 is highly flexible and exhibits high sequence variation compared with the other regions, the significance of which in diarrhoea induction remain unresolved. Here, it was shown that every amino acid substitution or deletion in the flexible C terminus resulted in altered conformation, multimerization, trypsin resistance and thioflavin T (ThT)binding, and affected DLP binding and the diarrhoea-inducing ability of the highly diarrhoeagenic SA11 and Hg18 $\triangle$N72 in suckling mice. These studies further revealed that high ThT fluorescence correlated with efficient diarrhoea induction, suggesting the importance of an optimal ThT-recognizable conformation in diarrhoea induction by purified NSP4. These results based on biological properties provide a possible conformational basis for understanding the influence of primary sequence variations on diarrhoea induction in newborn mice by purified NSP4s that cannot be explained by extensive sequence analyses