26,844 research outputs found
Conformational studies of various hemoglobins by natural-abundance 13C NMR spectroscopy
Studies of variously liganded hemoglobins (both from human and rabbit) by natural-abundance 13C NMR spectroscopy have revealed apparent conformational differences that have been interpreted on the basis of two quaternary structures for the α2ß2 tetramer, and variable tertiary structures for the individual α and ß subunits. In solution, rabbit hemoglobins appear to have somewhat more flexibility than human hemoglobins
Transient heat and mass transfer analysis of supercritical cryogenic storage systems with spherical static heaters Final report
Transient heat and mass transfer analysis of supercritical cryogenic storage systems with spherical static heaters by computer progra
Altering Enzymatic Activity: Recruitment of Carboxypeptidase Activity into an RTEM β-Lactamase/Penicillin-Binding Protein 5 Chimera
The D-Ala-D-Ala carboxypeptidases/transpeptidases (penicillin-binding proteins, PBPs) share considerable structural homology with class A β-lactamases (EC 3.5.2.6), although these β-lactamases have no observable D-Ala-D-Ala carboxypeptidase activity. With the objective of recruiting such activity into a β-lactamase background, we have prepared a chimeric protein by inserting a 28-amino acid segment of PBP-5 of Escherichia coli in place of the corresponding region of the RTEM-1 β-lactamase. The segment thus inserted encompasses two residues conserved in both families: Ser-70, which forms the acyl-enzyme intermediate during β-lactam hydrolysis, and Lys-73, whose presence has been shown to be necessary for catalysis. This chimera involves changes of 18 residues and gives a protein that differs at 7% of the residues from the parent. Whereas RTEM β-lactamase has no D-Ala-D-Ala carboxypeptidase activity, that of the chimera is significant and is, in fact, about 1% the activity of PBP-5 on diacetyl-L-Lys-D-Ala-D-Ala; in terms of free energy of activation, the chimera stabilizes the transition state for the reaction to within about 2.7 kcal/mol of the stabilization achieved by PBP-5. Furthermore, the chimera catalyzes hydrolysis exclusively at the carboxyl-terminal amide bond which is the site of cleavage by D-Ala-D-Ala carboxypeptidase. Though containing all those residues that are conserved throughout class A β-Lactamases and are thought to be essential for β-lactamase activity, the chimera has considerably reduced activity ({approx} 10^-5) on penams such as penicillins and ampicillins as substrates. As a catalyst, the chimera shows an induction period of {approx} 30 min, reflecting a slow conformational rearrangement from an inactive precursor to the active enzyme
Ionization behavior of the histidine residue in the catalytic triad of serine proteases
α-Lytic protease is a homologue of the mammalian serine proteases such as trypsin, chymotrypsin, and elastase, and its single histidine residue belongs to the Asp-His-Ser catalytic triad. This single histidine residue has been selectively enriched in the C-2 carbon with 13C. Magnetic resonance studies of the chemical shift and coupling constant (1Jch) behavior of this nucleus as a function of pH suggest that the imidazole ring is neutral above pH 5 and therefore that the group which is known to ionize with pKa near 6.7 must be the aspartic acid residue.
Implications of these new pKa assignments for the catalytic mechanism of serine proteases are discussed and include the absence of any need to separate charge during catalysis. The histidine residue plays two roles. (a) It insulates the aspartic acid from an aqueous environment and accordingly raises its pKa. (b) It serves as a bidentate base to accept a proton from the serine at one of its nitrogens and concertedly transfer a proton from its other nitrogen to the buried carboxylate anion during formation of the tetrahedral intermediate
The effect of parallel static and microwave electric fields on excited hydrogen atoms
Motivated by recent experiments we analyse the classical dynamics of a
hydrogen atom in parallel static and microwave electric fields. Using an
appropriate representation and averaging approximations we show that resonant
ionisation is controlled by a separatrix, and provide necessary conditions for
a dynamical resonance to affect the ionisation probability.
The position of the dynamical resonance is computed using a high-order
perturbation series, and estimate its radius of convergence. We show that the
position of the dynamical resonance does not coincide precisely with the
ionisation maxima, and that the field switch-on time can dramatically affect
the ionisation signal which, for long switch times, reflects the shape of an
incipient homoclinic. Similarly, the resonance ionisation time can reflect the
time-scale of the separatrix motion, which is therefore longer than
conventional static field Stark ionisation. We explain why these effects should
be observed in the quantum dynamics.
PACs: 32.80.Rm, 33.40.+f, 34.10.+x, 05.45.Ac, 05.45.MtComment: 47 pages, 20 figure
High-Potential C112D/M121X (X = M, E, H, L) Pseudomonas aeruginosa Azurins
Site-directed mutagenesis of Pseudomonas aeruginosa azurin
C112D at the M121 position has afforded a series of proteins with
elevated Cu^(II/I) reduction potentials relative to the CuII aquo ion.
The high potential and low axial hyperfine splitting (Cu^(II) electron
paramagnetic resonance A|) of the C112D/M121L protein are
remarkably similar to features normally associated with type 1
copper centers
Photometric study of southern SU UMa-type dwarf novae and candidates -- III: NSV 10934, MM Sco, AB Nor, CAL 86
We photometrically observed four southern dwarf novae in outburst (NSV 10934,
MM Sco, AB Nor and CAL 86). NSV 10934 was confirmed to be an SU UMa-type dwarf
nova with a mean superhump period of 0.07478(1) d. This star also showed
transient appearance of quasi-periodic oscillations (QPOs) during the final
growing stage of the superhumps. Combined with the recent theoretical
interpretation and with the rather unusual rapid terminal fading of normal
outbursts, NSV 10934 may be a candidate intermediate polar showing SU UMa-type
properties. The mean superhump periods of MM Sco and AB Nor were determined to
be 0.06136(4) d and 0.08438(2) d, respectively. We suggest that AB Nor belongs
to a rather rare class of long-period SU UMa-type dwarf novae with low
mass-transfer rates. We also observed an outburst of the suspected SU UMa-type
dwarf nova CAL 86. We identified this outburst as a normal outburst and
determined the mean decline rate of 1.1 mag/d.Comment: 13 pages, 23 figures, to appear in MNRAS. For more information, see
http://www.kusastro.kyoto-u.ac.jp/vsnet
Preliminary analysis of performance and loads data from the 2-megawatt mod-1 wind turbine generator
Preliminary test data on output power versus wind speed, rotor blade loads, system dynamic behavior, and start-stop characteristics on the Mod-1 wind turbine generator are presented. These data were analyzed statistically and are compared with design predictions of system performance and loads. To date, the Mod-1 wind turbine generator has produced up to 1.5 MW of power, with a measured power versus wind speed curve which agrees closely with design. Blade loads were measured at wind speeds up to 14 m/s and also during rapid shutdowns. Peak transient loads during the most severe shutdowns are less than the design limit loads. On the inboard blade sections, fatigue loads are approximately equal to the design cyclic loads. On the outboard blade sections, however, measured cyclic loads are significantly larger than design values, but they do not appear to exceed fatigue allowable loads as yet
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