Indeed, the region encompassing residues N33-F83 of BinB has been predicted to be α-helical in nature (Elangovan et al., 2000) and could be potentially required for the binding interaction or for the proper folding of the protein, a possibility highlighted by the lack of phenotype observed by three sets of mutations that targeted

motifs localized within this segment (32YNL34, 38SKK40, 52GYG54). Nevertheless, it seems clear that different elements within the protein’s N-terminal third are involved in forming a binding surface for the receptor. Further investigation of these elements, as well as the complementary binding region in the midgut receptor, will be carried out in order to provide data for developing strategies to improve the binary toxin’s insecticidal action on mosquito larvae. We thank the team from the insectarium for the technical support,

Christian Reis, Diogo PARP inhibitor Selleck Cyclopamine Chalegre, Lígia Ferreira and Maria da Conceição Costa for helpful discussion with the experimental procedures and the Program for Technological Development in Tools for Health PDTIS/FIOCRUZ for allowing the use of its facilities. This study was supported by the Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco-FACEPE (grant APQ 0427-2.13/08). Fig. S1. Sequence comparison of the BinA and BinB subunits of the binary toxin from Bacillus sphaericus strain 1593. Fig. S2. Immunoblotting of midgut microvilli proteins from Culex quinquefasciatus larvae bound RG7420 order to immobilized Bacillus sphaericus BinB proteins. Table S1. Forward oligonucleotides used for site-directed mutagenesis of the gene encoding the BinB subunit from Bacillus sphaericus

binary toxin. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Methods for in vivo monitoring of redox changes in different cellular compartments have been developed in recent years, and are mostly based on redox-sensitive variants of the green fluorescent protein (GFP). However, due to the thermodynamic stability of the introduced reactive disulfide bond, these sensors are limited to reducing compartments such as the cytosol and the mitochondria, and are not suited for more oxidizing environments such as the endoplasmic reticulum (ER). To overcome this problem, a family of redox-sensitive GFP variants that differed in their midpoint potential has been developed by the group of Remington (University of Oregon) and tested in vitro. Here, we report the first in vivo use of these novel roGFP1 variants for the measurement of redox conditions within the ER and cytosol in the yeast Pichia pastoris. With the fluorescence data obtained, it was possible to determine the reduction potential of the two compartments.