Anti-CGB monoclonal antibody, clone BCN514

Cytoglobin (Cgb) was discovered a decade ago and is a fourth member of the group of hexacoordinated globinfolded proteins. Although some crystal structures have been reported and several functions have been proposed for Cgb, its physiological role remains uncertain. In this study, we measured cyanide binding to the ferric state of the wild-type (WT) Cgb, and found that the binding consisted of multiple steps. These results indicated that Cgb may be comprised of several forms, and the presence of monomers, dimers, and tetramers was subsequently confirmed by SDS-PAGE. Remarkably, each species contained two distinguishable forms, and, in the monomer, analyses of alternative cysteine states suggested the presence of an intramolecular disulfide bond (monomer SS form) and a structure with unpaired thiol groups (monomer SH form). These confirmed that forms were separated by gel-exclusion chromatography, and that the cyanide binding of the separated fractions was again measured; they showed different affinities for cyanide, with the monomer fraction showing the highest affinity. In addition, the ferrous state in each fraction showed distinct carbon monoxide (CO)-binding properties, and the affinities for cyanide and CO suggested a linear correlation. Furthermore, we also prepared several variants involving the two cysteine residues. The C38S and C83S variants showed a binding affinity for cyanide similar to the value for the monomer SH form, and hence the fraction with the highest affinity for exogenous ligands was designated as a monomer SS form. We concluded that polymerization could be a mechanism that triggers the exertion of various physiological functions of this protein and that an appropriate disulfide bond between the two cysteine residues was critical for regulating the binding affinity of Cgb, which can act as a ROS scavenger, for exogenous ligands. (C) 2014 Elsevier Inc. All rights reserved.

We introduce a general approach for synthesizing multicomponent metal-decorated crumpled reduced graphene oxide nanocomposites using a one-step, continuous flame-based process. Crumpled reduced graphene oxide balls (CGB) were produced from graphene oxide (GO) in a High Temperature Reducing Jet (HTRJ) reactor. Moreover, CGBs were simultaneously decorated with different transition metal nanoparticles, including cobalt (Co), nickel (Ni), iron (Fe), and palladium (Pd). Various metal alloy-decorated crumpled reduced graphene oxide balls (M-CGBs) including CoPd-, CoNi-, CoPdNi-, and CoNiFe-CGBs were successfully synthesized using a general recipe. The key advantage of the HTRJ system over common flame-based aerosol synthesis methods is the separation of flame and product formation zones, which allows production and/or reduction of nanomaterials that can be reduced by H-2 in the presence of H2O. Nanomaterials are produced from aqueous precursors containing low-cost metal salts and dispersed GO. Electron microscopy and other characterization methods show the decoration of the CGBs with sub-4 nm diameter binary and ternary alloy, non-oxide transition metal nanoparticles of controlled compositions. The nanostructures made by this process can potentially be used as electrocatalysts for fuel cells, electrodes in batteries and supercapacitors, conductive inks for printed electronics, catalysts in wastewater treatment, and many other applications where a graphitized carbon-metal nanomaterial is needed.