Multiple cysteine residues are necessary for sorting and transport activity of the arsenite permease Acr3p from Saccharomyces cerevisiae

The yeast transporter Acr3p is a low affinity As(III)/H⁺ and Sb(III)/H⁺ antiporter located in the plasma membrane. It has been shown for bacterial Acr3 proteins that just a single cysteine residue, which is located in the middle of the fourth transmembrane region and conserved in all members of the Acr3 family, is essential for As(III) transport activity. Here, we report a systematic mutational analysis of all nine cysteine residues present in the Saccharomyces cerevisiae Acr3p. We found that mutagenesis of highly conserved Cys151 resulted in a complete loss of metalloid transport function. In addition, lack of Cys90 and Cys169, which are conserved in eukaryotic members of Acr3 family, impaired Acr3p trafficking to the plasma membrane and greatly reduced As(III) efflux, respectively. Mutagenesis of five other cysteines in Acr3p resulted in moderate reduction of As(III) transport capacities and sorting perturbations. Our data suggest that interaction of As(III) with multiple thiol groups in the yeast Acr3p may facilitate As(III) translocation across the plasma membrane.     

鹤顶兰胚囊发育过程中Ca~（2＋）分布的超微细胞化学定位（英文）

用焦锑酸盐沉淀法对鹤顶兰（Phaius tankervilliae）胚囊发育过程中的Ca2＋状态进行超微细胞化学定位。观察结果发现：功能大孢子时期,珠孔端的胚囊壁上开始出现小颗粒的Ca2＋沉淀,但功能大孢子细胞内未见明显的Ca2＋标记;四核胚囊时期胚囊壁上的Ca2＋沉淀明显增多,液泡膜上有Ca2＋沉淀出现,珠孔处的Ca2＋沉淀颗粒较大;成熟胚囊时期,胚囊壁上的Ca2＋沉淀进一步增多,且胚囊内Ca2＋分布明显增多,且极性明显,珠孔端助细胞、卵细胞比合点端反足细胞有更多的Ca2＋沉淀。鹤顶兰成熟胚囊内Ca2＋积累的来源有：（1）在胚囊成熟前主要由珠被细胞、珠细胞通过胞间连丝向胚囊运输;（2）以沉淀有大量Ca2＋的小泡形式跨过胚囊壁进入胚囊。     

水稻胞质雄性不育系珍汕97A及其保持系珍汕97B绒毡层发育过程中的Ca2+分布变化

利用焦锑酸钾沉淀法研究了野败不育系珍汕97A及其保持系珍汕97B绒毡层细胞的发育过程及其细胞中Ca2 的分布变化。研究发现保持系绒毡层细胞在单核花粉晚期才开始迅速解体,而不育系绒毡层细胞在花粉母细胞时期就开始出现核膜、细胞膜解体,此过程持续到二核花粉时期。珍汕97A绒毡层细胞从花粉母细胞时期开始,细胞质内有少量颗粒状的Ca2 沉淀;减数分裂时期,绒毡层细胞的内切向壁表面有大量大颗粒的Ca2 沉淀;单核花粉时期绒毡层细胞周围集聚一层Ca2 沉淀。而保持系绒毡层细胞遮花粉母细胞时期和减数分裂时期细胞内没有Ca2 沉淀;单核花粉时期绒毡层细胞内的Ca2 沉淀主要分布在解体的细胞质内。推测绒毡层细胞结构发育的异常和Ca2 的异常分布可能与花粉的败育有关。     

Role of conserved histidine residues in metalloactivation of the ArsA ATPase

The ArsA ATPase is the catalytic subunit of a pump that is responsible for resistance to arsenicals and antimonials in Escherichia coli. Arsenite or antimonite allosterically activates the ArsA ATPase activity. ArsA homologues from eubacteria, archaea and eukarya have a signature sequence (DTAPTGHT) that includes a conserved histidine. The ArsA ATPase has two such conserved motifs, one in the NH₂-terminal (A1) half and the other in the COOH-terminal (A2) half of the protein. These sequences have been proposed to be signal transduction domains that transmit the information of metal occupancy at the allosteric to the catalytic site to activate ATP hydrolysis. The role of the conserved residues His148 and His453, which reside in the A1 and A2 signal transduction domains respectively, was investigated by mutagenesis to create H148A, H453A or H148A/H453A ArsAs. Each altered protein exhibited a decrease in the V _max of metalloid-activated ATP hydrolysis, in the order wild type ArsA>H148A>H453A>H148A/H453A. These results suggest that the histidine residues play a role in transmission of the signal between the catalytic and allosteric sites.     

Photoluminescence properties of a new orange–red‐emitting Sm3+–La3SbO7 phosphor

The antimonate compound La₃SbO₇ has high chemical stability, lattice stiffness and thermal stability. Orange–red‐emitting antimonate‐based phosphors La₃SbO₇:xSm³⁺ (x = 0.02, 0.05, 0.08, 0.10, 0.15, 0.20 and 0.25) were synthesized. The phase structure and photoluminescence properties of these phosphors were investigated. The emission spectrum obtained on excitation at 407 nm contained exclusively the characteristic emissions of Sm³⁺ at 568, 608, 654 and 716 nm, which correspond to the transitions from ⁴G_5/2 to ⁶H_5/2, ⁶H_7/2, ⁶H_9/2 and ⁶H_11/2 of Sm³⁺, respectively. The strongest emission was located at 608 nm due to the ⁴G_5/2→⁶H_7/2 transition of Sm³⁺, generating bright orange–red light. The critical quenching concentration of Sm³⁺ in La₃SbO₇:Sm³⁺ phosphor was determined as 10% and the energy transfer between Sm³⁺ was found to be through an exchange interaction. The International Commission on Illumination chromaticity coordinates of the La₃SbO₇:0.10Sm³⁺ phosphors are located in the orange–red region. The La₃SbO₇:Sm³⁺ phosphors may be potentially used as red phosphors for white light‐emitting diodes. Copyright © 2015 John Wiley & Sons, Ltd.