Structure of S. aureus HPPK and discovery of a new inhibitor

The first structural and biophysical data on the folate pathway enzyme and drug target, 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK), from the pathogenic bacterium Staphylococcus aureus is presented. HPPK is the second essential enzyme in the folate biosynthesis pathway, responsible for catalysing pyrophosphoryl transfer from cofactor (ATP) to the substrate (6-hydroxymethyl- 7,8-dihydropterin, HMDP). In-silico screening led to the discovery of a substrate competitive inhibitor, San1, which was subsequently co-crystallised with HPPK. A 1.65 Å resolution x-ray structure showed this to bind at the pterin site sharing many of the key intermolecular interactions of the substrate. ITC and SPR measurements yielded an equilibrium binding constant, Kd, of ~13 μM for San1. An IC50 of ~12 μM was determined by means of a new convenient tri-enzyme-coupled spectrophotometric assay. ITC and SPR further showed that the San1 inhibitor has no requirement for magnesium or ATP cofactor for competitive binding to the substrate site. According to 15N heteronuclear NMR measurements, the fast motion of the pterin loop (L2) is partially dampened in the ternary complex between SaHPPK, HMDP and , -methylene adenosine 5-triphosphate (AMPCPP), but the ATP loop (L3) remains mobile on the μs timescale. In contrast, for the SaHPPK/San1/AMPCPP ternary complex, loop L2 becomes rigid on the fast timescale and loop L3 becomes more ordered which are supported by a large entropic penalty associated with San1 binding as revealed by ITC. Backbone assignments and chemical shift perturbations implicate the sulphur in San1 as a likely important loop L2/L3 stabilizing mediato

ROTSE All Sky Surveys for Variable Stars I: Test Fields

The ROTSE-I experiment has generated CCD photometry for the entire Northern sky in two epochs nightly since March 1998. These sky patrol data are a powerful resource for studies of astrophysical transients. As a demonstration project, we present first results of a search for periodic variable stars derived from ROTSE-I observations. Variable identification, period determination, and type classification are conducted via automatic algorithms. In a set of nine ROTSE-I sky patrol fields covering about 2000 square degrees we identify 1781 periodic variable stars with mean magnitudes between m_v=10.0 and m_v=15.5. About 90% of these objects are newly identified as variable. Examples of many familiar types are presented. All classifications for this study have been manually confirmed. The selection criteria for this analysis have been conservatively defined, and are known to be biased against some variable classes. This preliminary study includes only 5.6% of the total ROTSE-I sky coverage, suggesting that the full ROTSE-I variable catalog will include more than 32,000 periodic variable stars.Comment: Accepted for publication in AJ 4/00. LaTeX manuscript. (28 pages, 11 postscript figures and 1 gif

The RR Lyrae Distance Scale

We review seven methods of measuring the absolute magnitude M_V of RR Lyrae stars in light of the Hipparcos mission and other recent developments. We focus on identifying possible systematic errors and rank the methods by relative immunity to such errors. For the three most robust methods, statistical parallax, trigonometric parallax, and cluster kinematics, we find M_V (at [Fe/H] = -1.6) of 0.77 +/- 0.13, 0.71 +/- 0.15, 0.67 +/- 0.10. These methods cluster consistently around 0.71 +/- 0.07. We find that Baade-Wesselink and theoretical models both yield a broad range of possible values (0.45-0.70 and 0.45-0.65) due to systematic uncertainties in the temperature scale and input physics. Main-sequence fitting gives a much brighter M_V = 0.45 +/- 0.04 but this may be due to a difference in the metallicity scales of the cluster giants and the calibrating subdwarfs. White-dwarf cooling-sequence fitting gives 0.67 +/- 0.13 and is potentially very robust, but at present is too new to be fully tested for systematics. If the three most robust methods are combined with Walker's mean measurement for 6 LMC clusters, V_{0,LMC} = 18.98 +/- 0.03 at [Fe/H] = -1.9, then mu_{LMC} = 18.33 +/- 0.08.Comment: Invited review article to appear in: `Post-Hipparcos Cosmic Candles', A. Heck & F. Caputo (Eds), Kluwer Academic Publ., Dordrecht, in press. 21 pages including 1 table; uses Kluwer's crckapb.sty LaTeX style file, enclose

CU Comae: a new field double-mode RR Lyrae, the most metal poor discovered to date

We report the discovery of a new double-mode RR Lyrae variable (RRd) in the field of our Galaxy: CU Comae. CU Comae is the sixth such RRd identified to date and is the most metal-poor RRd ever detected. Based on BVI CCD photometry spanning eleven years of observations, we find that CU Comae has periods P0=0.5441641 +/-0.0000049d and P1=0.4057605 +/-0.0000018d. The amplitude of the primary (first-overtone) period of CU Comae is about twice the amplitude of the secondary (fundamental) period. The combination of the fundamental period of pulsation P0 and the period ratio of P1/P0=0.7457 places the variable on the metal-poor side of the Petersen diagram, in the region occupied by M68 and M15 RRd's. A mass of 0.83 solar masses is estimated for CU Comae using an updated theoretical calibration of the Petersen diagram. High resolution spectroscopy (R=30,000) covering the full pulsation cycle of CU Comae was obtained with the 2.7 m telescope of the Mc Donald Observatory, and has been used to build up the radial velocity curve of the variable. Abundance analysis done on the four spectra taken near minimum light (phase: 0.54 -- 0.71) confirms the metal poor nature of CU Comae, for which we derive [Fe/H]=-2.38 +/-0.20. This value places this new RRd at the extreme metal-poor edge of the metallicity distribution of the RR Lyrae variables in our Galaxy.Comment: 21 pages including 8 Tables, Latex, 11 Figures. Accepted for publication in The Astronomical Journal, October 2000 issu

Distances and ages of globular clusters using Hipparcos parallaxes of local subdwarfs

We discuss the impact of Population II and Globular Cluster (GCs) stars on the derivation of the age of the Universe, and on the study of the formation and early evolution of galaxies, our own in particular. The long-standing problem of the actual distance scale to Population II stars and GCs is addressed, and a variety of different methods commonly used to derive distances to Population II stars are briefly reviewed. Emphasis is given to the discussion of distances and ages for GCs derived using Hipparcos parallaxes of local subdwarfs. Results obtained by different authors are slightly different, depending on different assumptions about metallicity scale, reddenings, and corrections for undetected binaries. These and other uncertainties present in the method are discussed. Finally, we outline progress expected in the near future.Comment: Invited review article to appear in: `Post-Hipparcos Cosmic Candles', A. Heck & F. Caputo (Eds), Kluwer Academic Publ., Dordrecht, in press. 22 pages including 3 tables and 2 postscript figures, uses Kluwer's crckapb.sty LaTeX style file, enclose

The OSACA Database and a Kinematic Analysis of Stars in the Solar Neighborhood

We transformed radial velocities compiled from more than 1400 published sources, including the Geneva--Copenhagen survey of the solar neighborhood (CORAVEL-CfA), into a uniform system based on the radial velocities of 854 standard stars in our list. This enabled us to calculate the average weighted radial velocities for more than 25~000 HIPPARCOS stars located in the local Galactic spiral arm (Orion arm) with a median error of +-1 km/s. We use these radial velocities together with the stars' coordinates, parallaxes, and proper motions to determine their Galactic coordinates and space velocities. These quantities, along with other parameters of the stars, are available from the continuously updated Orion Spiral Arm CAtalogue (OSACA) and the associated database. We perform a kinematic analysis of the stars by applying an Ogorodnikov-Milne model to the OSACA data. The kinematics of the nearest single and multiple main-sequence stars differ substantially. We used distant (r\approx 0.2 kpc) stars of mixed spectral composition to estimate the angular velocity of the Galactic rotation -25.7+-1.2 km/s/kpc, and the vertex deviation,l=13+-2 degrees, and detect a negative K effect. This negative K effect is most conspicuous in the motion of A0-A5 giants, and is equal to K=-13.1+-2.0 km/s/kpc.Comment: 16 pages, 8 figure

Astrometry with Hubble Space Telescope: A Parallax of the Fundamental Distance Calibrator RR Lyrae

We present an absolute parallax and relative proper motion for the fundamental distance scale calibrator, RR Lyr. We obtain these with astrometric data from FGS 3, a white-light interferometer on HST. We find $\pi_{abs} = 3.82 \pm 0.2$ mas. Spectral classifications and VRIJHKT$_2$M and DDO51 photometry of the astrometric reference frame surrounding RR Lyr indicate that field extinction is low along this line of sight. We estimate =0.07\pm0.03 for these reference stars. The extinction suffered by RR Lyr becomes one of the dominant contributors to the uncertainty in its absolute magnitude. Adopting the average field absorption, =0.07 \pm 0.03, we obtain M_V^{RR} = 0.61 ^{-0.11}_{+0.10}. This provides a distance modulus for the LMC, m-M = 18.38 - 18.53^{-0.11}_{+0.10} with the average extinction-corrected magnitude of RR Lyr variables in the LMC, , remaining a significant uncertainty. We compare this result to more than 80 other determinations of the distance modulus of the LMC.Comment: Several typos corrected. To appear in The Astronomical Journal, January 200