Structures of the multidrug exporter AcrB reveal a proximal multisite drug-binding pocket

AcrB and its homologues are the principal multidrug transporters in Gram-negative bacteria and are important in antibiotic drug tolerance. AcrB is a homotrimer that acts as a tripartite complex with the outer membrane channel TolC and the membrane fusion protein AcrA. Minocycline and doxorubicin have been shown to bind to the phenylalanine cluster region of the binding monomer. Here we report the crystal structures of AcrB bound to the high-molecular-mass drugs rifampicin and erythromycin. These drugs bind to the access monomer, and the binding sites are located in the proximal multisite binding pocket, which is separated from the phenylalanine cluster region (distal pocket) by the Phe-617 loop. Our structures indicate that there are two discrete multisite binding pockets along the intramolecular channel. High-molecular-mass drugs first bind to the proximal pocket in the access state and are then forced into the distal pocket in the binding state by a peristaltic mechanism involving subdomain movements that include a shift of the Phe-617 loop. By contrast, low-molecular-mass drugs, such as minocycline and doxorubicin, travel through the proximal pocket without specific binding and immediately bind to the distal pocket. The presence of two discrete, high-volume multisite binding pockets contributes to the remarkably broad substrate recognition of AcrB.     

CXorf6 is a causative gene for hypospadias

46,XY disorders of sex development (DSD) refer to a wide range of abnormal genitalia, including hypospadias, which affects approximately 0.5% of male newborns. We identified three different nonsense mutations of CXorf6 in individuals with hypospadias and found that its mouse homolog was specifically expressed in fetal Sertoli and Leydig cells around the critical period for sex development. These data imply that CXorf6 is a causative gene for hypospadias.     

Chiral influence of sex pheromonal substances on responses of the male American cockroach

Summary In order to elucidate the roles of optical enantiomers of sex pheromonal substances of the American cockroach, behavioral assays with a single enantiomer and with mixtures of enantiomers of sex pheromone mimics were carried out. Inactive enantiomers [(–)-enantiomers] had no influence on the potency of enantiomers active as sex pheromones [(+)-enantiomers]. By analysis of the results from EAG recordings with single and mixed sample of the enantiomers, it was confirmed that (–)-enantiomers did not react with the sex pheromone receptors which are responsive to (+)-enantiomers.We acknowledge technical assistance from Miss R. Kimura.     

Insulin-like effect of dichloroacetic acid on hexose transport in Swiss 3T3 cells

Summary Hexose transport in Swiss 3T3 cells was increased by treatment with dichloroacetic acid as well as by treatment with insulin. Neither extra-nor intracellular Ca²⁺ was found to be involved in their stimulatory action. On the other hand, the removal of intracellular Mg²⁺ resulted in a loss of the stimulation. These results suggest that dichloroacetic acid stimulates the hexose transport in Mg²⁺-dependent manner, similar to that of insulin.This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture, and the Ministry of Health and Welfare of Japan.     

Subtypes of protein kinase C in rat cerebral microvessels

Summary Protein kinase C in rat cerebral microvessels was characterized. By hydroxyapatite column chromatography, protein kinase C in the soluble fraction was resolved into two major peaks corresponding to type II and III enzymes, in the proportions of 57% and 38%, respectively. Since each subtype is considered to have a distinct role, the high proportion of type II enzyme found in this study suggests that this type may be involved in specific functions of the cerebral microvessels.     

Possible involvement of protein kinase C in the stimulation of amino acid transport by phorbol ester,platelet-derived growth factor and A23187 in Swiss 3T3 cells

Summary Stimulation of amino acid transport induced by phorbol-12, 13-dibutyrate, platelet-derived growth factor or A23187 was not observed in cells lacking protein kinase C. On the other hand, stimulation of transport by epidermal growth factor or insulin was not affected. These results suggested that the stimulation of amino acid transport is mediated by at least two separate pathways.This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture, and the Ministry of Health and Welfare of Japan.     

Possible involvement of protein kinase C in the stimulation of amino acid transport by phorbol ester, platelet-derived growth factor and A23187 in Swiss 3T3 cells

Stimulation of amino acid transport induced by phorbol-12,13-dibutyrate, platelet-derived growth factor or A23187 was not observed in cells lacking protein kinase C. On the other hand, stimulation of transport by epidermal growth factor or insulin was not affected. These results suggested that the stimulation of amino acid transport is mediated by at least two separate pathways.     

Self-regeneration of a Pd-perovskite catalyst for automotive emissions control

Catalytic converters are widely used to reduce the amounts of nitrogen oxides, carbon monoxide and unburned hydrocarbons in automotive emissions. The catalysts are finely divided precious-metal particles dispersed on a solid support. During vehicle use, the converter is exposed to heat, which causes the metal particles to agglomerate and grow, and their overall surface area to decrease. As a result, catalyst activity deteriorates. The problem has been exacerbated in recent years by the trend to install catalytic converters closer to the engine, which ensures immediate activation of the catalyst on engine start-up, but also places demanding requirements on the catalyst's heat resistance. Conventional catalyst systems thus incorporate a sufficient excess of precious metal to guarantee continuous catalytic activity for vehicle use over 50,000 miles (80,000 km). Here we use X-ray diffraction and absorption to show that LaFe(0.57)Co(0.38)Pd(0.05)O(3), one of the perovskite-based catalysts investigated for catalytic converter applications since the early 1970s, retains its high metal dispersion owing to structural responses to the fluctuations in exhaust-gas composition that occur in state-of-the-art petrol engines. We find that as the catalyst is cycled between oxidative and reductive atmospheres typically encountered in exhaust gas, palladium (Pd) reversibly moves into and out of the perovskite lattice. This movement appears to suppress the growth of metallic Pd particles, and hence explains the retention of high catalyst activity during long-term use and ageing.