The Factory and the Beehive. II. Activity and Rotation is Praesepe and the Hyades

Open clusters are collections of stars with a single, well-determined age, and can be used to investigate the connections between angular-momentum evolution and magnetic activity over a star\u27s lifetime. We present the results of a comparative study of the relationship between stellar rotation and activity in two benchmark open clusters: Praesepe and the Hyades. As they have the same age and roughly solar metallicity, these clusters serve as an ideal laboratory for testing the agreement between theoretical and empirical rotation-activity relations at 600 Myr. We have compiled a sample of 720 spectra—more than half of which are new observations—for 516 high-confidence members of Praesepe; we have also obtained 139 new spectra for 130 high-confidence Hyads. We have also collected rotation periods (P rot) for 135 Praesepe members and 87 Hyads. To compare Hα emission, an indicator of chromospheric activity, as a function of color, mass, and Rossby number Ro , we first calculate an expanded set of χ values, with which we can obtain the Hα to bolometric luminosity ratio, L Hα/L bol, even when spectra are not flux-calibrated and/or stars lack reliable distances. Our χ values cover a broader range of stellar masses and colors (roughly equivalent to spectral types from K0 to M9), and exhibit better agreement between independent calculations, than existing values. Unlike previous authors, we find no difference between the two clusters in their Hα equivalent width or L Hα/L bol distributions, and therefore take the merged Hα and P rot data to be representative of 600 Myr old stars. Our analysis shows that Hα activity in these stars is saturated for . Above that value activity declines as a power-law with slope , before dropping off rapidly at Ro 0.4. These data provide a useful anchor for calibrating the age-activity-rotation relation beyond 600 Myr

Rapid and sensitive identification of major histocompatibility complex class I-associated tumor peptides by Nano-LC MALDI MS/MS

Identification of major histocompatibility complex (MHC)-associated peptides recognized by T-lymphocytes is a crucial prerequisite for the detection and manipulation of specific immune responses in cancer, viral infections, and autoimmune diseases. Unfortunately immunogenic peptides are less abundant species present in highly complex mixtures of MHC-extracted material. Most peptide identification strategies use microcapillary LC coupled to nano-ESI MS/MS in a challenging on-line approach. Alternatively MALDI PSD analysis has been applied for this purpose. We report here on the first off-line combination of nanoscale (nano) LC and MALDI TOF/TOF MS/MS for the identification of naturally processed MHC peptide ligands. These peptides were acid-eluted from human leukocyte antigen (HLA)-A2, HLA-A3, and HLA-B/-C complexes separately isolated from a renal cell carcinoma cell lysate using HLA allele-specific antibodies. After reversed-phase HPLC, peptides were further fractionated via nano-LC. This additional separation step provided a substantial increase in the number of detectable candidate species within the complex peptide pools. MALDI MS/MS analysis on nano-LC-separated material was then sufficiently sensitive to rapidly identify more than 30 novel HLA-presented peptide ligands. Peptide sequences contained perfect anchor amino acid residues described previously for HLA-A2, HLA-A3, and HLA-B7. The most promising candidate for a T-cell epitope is an HLA-B7-binding nonamer peptide derived from the tumor-associated gene NY-BR-16. To demonstrate the sensitivity of our approach we characterized peptides binding to HLA-C molecules that are usually expressed at the cell surface at approximately only 10% the levels of HLA-A or HLA-B. In fact, multiple renal cell carcinoma peptides were identified that contained anchor amino acid residues of HLA-Cw5 and HLA-Cw7. We conclude that the nano-LC MALDI MS/MS approach is a sensitive tool for the rapid and automated identification of MHC-associated tumor peptides.Sandra Hofmann, Matthias Glückmann, Sandra Kausche, Andrea Schmidt, Carsten Corvey, Rudolf Lichtenfels, Christoph Huber, Christian Albrecht, Michael Karas and Wolfgang Her

A four-subunit cytochrome bc1 complex complements the respiratory chain of Thermus thermophilus

Several components of the respiratory chain of the eubacterium Thermus thermophilus have previously been characterized to various extent, while no conclusive evidence for a cytochrome bc(1) complex has been obtained. Here, we show that four consecutive genes encoding cytochrome bc(1) subunits are organized in an operon-like structure termed fbcCXFB. The four gene products are identified as genuine subunits of a cytochrome bc(1) complex isolated from membranes of T. thermophilus. While both the cytochrome b and the FeS subunit show typical features of canonical subunits of this respiratory complex, a further membrane-integral component (FbcX) of so far unknown function copurifies as a subunit of this complex. The cytochrome c(1) carries an extensive N-terminal hydrophilic domain, followed by a hydrophobic, presumably membrane-embedded helical region and a typical heme c binding domain. This latter sequence has been expressed in Escherichia coli, and in vitro shown to be a kinetically competent electron donor to cytochrome c(552), mediating electron transfer to the ba(3) oxidase. Identification of this cytochrome bc(1) complex bridges the gap between the previously reported NADH oxidation activities and terminal oxidases, thus, defining all components of a minimal, mitochondrial-type electron transfer chain in this evolutionary ancient thermophile