Budding yeast Cdc13-Stn1-Ten1 (CST) complex plays an essential role in telomere protection and maintenance, and has been proposed to be a telomere-specific replication protein A (RPA)-like complex. Previous genetic and structural studies revealed a close resemblance between Stn1-Ten1 and RPA32-RPA14. However, the relationship between Cdc13 and RPA70, the largest subunit of RPA, has remained unclear. Here, we report the crystal structure of the N-terminal OB (oligonucleotide/oligosaccharide binding) fold of Cdc13. Although Cdc13 has an RPA70-like domain organization, the structures of Cdc13 OB folds are significantly different from their counterparts in RPA70, suggesting that they have distinct evolutionary origins. Furthermore, our structural and biochemical analyses revealed unexpected dimerization by the N-terminal OB fold and showed that homodimerization is probably a conserved feature of all Cdc13 proteins. We also uncovered the structural basis of the interaction between the Cdc13 N-terminal OB fold and the catalytic subunit of DNA polymerase α (Pol1), and demonstrated a role for Cdc13 dimerization in Pol1 binding. Analysis of the phenotypes of mutants defective in Cdc13 dimerization and Cdc13-Pol1 interaction revealed multiple mechanisms by which dimerization regulates telomere lengths in vivo. Collectively, our findings provide novel insights into the mechanisms and evolution of Cdc13.

We have investigated six nanomaterials for their applicability as surfaces for the analyses of peptides and proteins using surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS). Gold nanoparticles (NPs) were useful nanomaterials for small analytes (e.g., glutathione); Pt nanosponges and Fe 3 O 4 NPs were efficient nanomaterials for proteins, with an upper detectable mass limit of ca. 25 kDa. Nanomaterials have several advantages over organic matrices, including lower limits of detection for small analytes and lower batch-to-batch variations (fewer problems associated with "sweet spots"), when used in laser desorption/ionization mass spectrometry. (J Am Soc Mass Spectrom 2010, 21, 1204 -1207) © 2010 American Society for Mass Spectrometry S urface-assisted laser desorption/ionization mass spectrometry (SALDI-MS) was developed recently using nanomaterials rather than organic compounds as matrices, for the determination of analytes of interest. For example, laser desorption/ionization (LDI) of intact proteins and protein aggregates in the presence of glycerol has been demonstrated using cobalt particles (ca. 30 nm) [1]. Similar to the role played by organic matrices, the particles absorb energy from the laser irradiation and transfer it efficiently to the analytes, thereby inducing desorption and ionization. Mixtures of graphite particles (2-150 m) and glycerol have been employed in the analysis of proteins and peptides [2,3]. Several other nanomaterials, including carbon nanotubes, nanodiamonds, and various nanoparticles (NPs, namely SiO 2 , ZnS, TiO 2 , Fe 3 O 4 , Fe 3 O 4 /TiO 2 , and Au) are also useful-without the addition of glycerol-for SALDI-MS [4 -12]. Because of their unique chemical and physical properties, NPs can also act as selective probes and/or efficient ionization nanomaterials. For example, Au and TiO 2 NPs are suitable for the concentration and ionization of aminothiols and catechins, respectively, in SALDI-MS [8,11]. One other advantage of using NPs is that fewer "sweet spots" are formed, thereby maximizing reproducibility. Although NPs have been used successfully for the determination of a range of analytes (from small analytes to proteins), a review of the literature reveals that the various NPs provide quite different results in terms of sensitivity, reproducibility, and mass range. Thus, our aim in this study was to evaluate the performance of several types of NPs for the analysis of peptides and proteins. ExperimentalSix nanomaterials-Au NPs, TiO 2 NPs, Se NPs, CdTe quantum dots (QDs), Fe 3 O 4 NPs, and Pt nanosponges (NSPs)-were tested for the SALDI-MS-based analyses of peptides and proteins; they were prepared in aqueous solutions and characterized according to procedures described in the literature [8,11,[13][14][15][16]. A twolayer preparation method was applied to deposit the nanomaterials and samples onto the metal plates used in SALDI-MS. First, one of the nanomaterial solutions (1 L) was deposited into one of the wells of the MS plate and dried under ambient cond...

This study presents a new magnetic bead-based microfluidic platform, which integrates three major modules for rapid leukocytes purification, genomic DNA (gDNA) extraction and fast analysis of genetic gene. By utilizing microfluidic technologies and magnetic beads conjugated with CD 15/45 antibodies, leukocytes in a human whole blood sample can be first purified and concentrated, followed by extraction of gDNA utilizing surface-charge switchable, DNA-specific, magnetic beads in the lysis solution. Then, specific genes associated with genetic diseases can be amplified by an on-chip polymerase chain reaction (PCR) process automatically. The whole pretreatment process including the leukocytes purification and gDNA extraction can be performed in an automatic fashion with the incorporation of the built bio-separators consisting of microcoils array within less than 20 min. The detection of single nucleotide polymorphism (SNP) genotyping of methylenetetra-hydrofolate reductase (MTHFR) C677T region associated with an increased risk of genetic diseases was further performed to demonstrate the capability of the proposed system. The extracted gDNA can be transported into a micro PCR chamber for on-chip fast nucleic acid amplification of detection genes with minimum human intervention. Hence, the developed system may provide a powerful automated platform for pretreatment of human leukocytes, gDNA extraction and fast analysis of genetic gene.

MnO 2 has been considered as the most promising bifunctional electrocatalyst toward oxygen reduction reaction (ORR) and oxygen evolution reaction (OER). Despite their highly active ORR performance, the OER catalytic activity of MnO 2 species is still far from satisfying. Herein, for the first time, highly active OER catalytic NiFe layered double hydroxides (NiFe LDHs) are combined with MnO 2 via a selective electrodeposition method to form a Janus electrode in which the MnO 2 and NiFe LDHs are in situ grown on two sides of a porous nickel foam (MnO 2 -NiFe/Ni). The MnO 2 -NiFe/Ni electrode exhibits excellent bifunctional catalytic activity and stability for both ORR and OER compared to bare MnO 2 on account of the rational design of the Janus bifunctional configuration separating OER and ORR active materials. Moreover, such a Janus MnO 2 -NiFe air electrode endows the zinc−air battery with better cycling stability and energy efficiency than the bare MnO 2 electrode. Our work demonstrates a novel Janus electrode configuration to design high-performance electrocatalysts for energy storage and conversion applications.