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Ambiguities in amino acid sequences are a potential problem in X-ray crystallographic studies of proteins. Amino acid side chains often cannot be reliably identified from the electron density. Many protein crystal structures that are now being solved are simple variants of a known wild-type structure. Thus, cloning artifacts or other untoward events can readily lead to cases in which the proposed sequence is not correct. An example is presented showing that mass spectrometry provides an excellent tool for analyzing suspected errors. The X-ray crystal structure of an insertion mutant of Staphylococcal nuclease has been solved to 1.67 Å resolution and refined to a crystallographic R value of 0.170 [Keefe & Lattman (1992). In preparation]. A single residue has been inserted in the C-terminal α helix. The inserted amino acid was believed to be an alanine residue, but the final electron density maps strongly indicated that a glycine had been inserted instead. To confirm the observations from the X-ray data, matrix-assisted laser desorption mass spectrometry was employed to verify the glycine insertion. This mass spectrometric technique has sufficient mass accuracy to detect the methyl group that distinguishes glycine from alanine and can be extended to the more common situation in which crystallographic measurements suggest a problem with the sequence, but cannot pinpoint its location or nature.  相似文献   
84.
The skew angle causes a discrepancy in determining the reader-to-writer offset (RWO) when using different periodical patterns in track profile tests. It also separates the peak overwrite (OW) from the peak high frequency amplitude HFA, (1 T periodical pattern) on corresponding track profiles. Furthermore, higher track density and larger skew angle exacerbate the skew effect and induce more RWO error, thus impacting the parametric performance optimization. Simulation studies are used to interpret the skew effect on the RWO determination and OW cross-track characteristics. Based on experimental investigations and simulation analyses, using the HFA, track profile for deriving the optimal RWO is proposed for spin-stand tests. Actual parametric characterization has proven that the optimal RWO minimized the skew effect and the RWO error, thus improving the parametric performance and reducing the test variation. The method is beneficial and necessary for the high track density characterization.  相似文献   
85.
    
A new 11 amino acid linear peptide named roseabol A (1) and the known compound 13-oxo-trans-9,10-epoxy-11(E)-octadecenoic acid (2) were isolated from the fungus Clonostachys rosea. Combined NMR and MS analysis revealed that roseabol A (1) contained amino acid residues characteristic of the peptaibol family of peptides such as isovaline, α-aminoisobutyric acid, hydroxyproline, leucinol, and an N-terminal isovaleric acid moiety. The amino acid sequence was established by a combination of NMR studies and tandem MS fragmentation analyses, and the absolute configurations of the constituent amino acids of 1 were determined by the advanced Marfey’s method. Compound 2 showed inhibitory activity against Merkel cell carcinoma, a rare and difficult-to-treat type of skin cancer, with an IC50 value of 16.5 μM.  相似文献   
86.
The size (volume) of a large tetrametallic molecular square, that has resisted characterization by mass spectrometry, has been determined by pulsed-field gradient NMR spectroscopy, a technique that reports on self-diffusion coefficients. These scale inversely with hydrodynamic radii, which in turn scale approximately as the cube of the assembly's mass. The technique has also been used to determine whether NMR spectral complexities observed for the new compound are due to contamination with chemically related assemblies, or instead reflect the intrinsic structural complexities of the compound itself.  相似文献   
87.
A frequency-domain based system for measuring acoustic impedance and reflection coefficient is described. The calibration procedure uses a least-mean-squares approximation to the Thevenin parameters describing the source and receiver characteristics in which the data measured on closed, cylindrical tubes are matched to a viscothermal tube model. The system is intended for use in acoustical measurement in human ear canals, in which the cross-sectional area of the ear canal at the point of insertion is imprecisely known. This area is acoustically estimated from the impedance data, and the reflection coefficient is calculated in terms of this area and the impedance data. Measurements on a variety of closed tubes show the method is accurate over the frequency range investigated (less than 10.7 kHz). The time-domain reflection function is evaluated by transforming the reflection coefficient from the frequency domain, but the finite bandwidth of the measured data limits the accuracy of time-domain response measurements. The method is well suited for frequency-domain measurements in human ear canals.  相似文献   
88.
The Kramers-Kronig transformation has been extensively applied in optical spectroscopy to calculate the real component of an optical quantity from the imaginary component, such as the real refractive index from the imaginary component, or vice versa. In this paper, the traditional proof of the Kramers-Kronig transformation, and its application to the complex refractive index, complex dielectric constant, and complex molar polarizability, are reviewed. Often the imaginary components of these quantities are fitted with standard lineshapes such as the Gaussian, Lorentzian, or Classical Damped Harmonic Oscillator (CDHO) lineshapes. It is shown that the usual Gaussian and Lorentzian lineshapes do not meet the physical criteria of these imaginary components nor the conditions of the Kramers-Kronig transformation since they are not odd functions of wavenumber. However, the CDHO lineshape meets the physical criteria of the imaginary components of these optical quantities and the Kramers-Kronig transformation. Modifications are presented that make the Gaussian and Lorentzian odd. The Gaussian decays so fast that the modification is not needed in practice; however, the Lorentzian is much slower to decay and thus modification is necessary whenever fitting peaks below approximately 250 cm(-1). Since the computational difference between the usual Lorentzian and modified Lorentzian is negligible, the author recommends that only the modified Lorentzian be used when fitting bands with a Lorentzian lineshape. Copyright 2001 Academic Press.  相似文献   
89.
This study evaluated the accuracy of acoustic response tests in predicting conductive hearing loss in 161 ears of subjects from the age of 2 to 10 yr, using as a "gold standard" the air-bone gap to classify ears as normal or impaired. The acoustic tests included tympanometric peak-compensated static admittance magnitude (SA) and tympanometric gradient at 226 Hz, and admittance-reflectance (YR) measurements from 0.5 to 8 kHz. The performance of individual, frequency-specific, YR test variables as predictors was assessed. By applying logistic regression (LR) and discriminant analysis (DA) techniques to the multivariate YR response, two univariate functions were calculated as the linear combinations of YR variables across frequency that best separated normal and impaired ears. The tympanometric and YR tests were also combined in a multivariate manner to test whether predictive efficacy improved when 226-Hz tympanometry was added to the predictor set. Conductive hearing loss was predicted based on air-bone gap thresholds at 0.5 and 2 kHz, and on a maximum air-bone gap at any octave frequency from 0.5 to 4 kHz. Each air-bone gap threshold ranged from 5 to 30 dB in 5-dB steps. Areas under the relative operating characteristic curve for DA and LR were larger than for reflectance at 2 kHz, SA and Gr. For constant hit rates of 80% and 90%, both DA and LR scores had lower false-alarm rates than tympanometric tests-LR achieved a false-alarm rate of 6% for a sensitivity of 90%. In general, LR outperformed DA as the multivariate technique of choice. In predicting an impairment at 0.5 kHz, the reflectance scores at 0.5 kHz were less accurate predictors than reflectance at 2 and 4 kHz. This supports the hypothesis that the 2-4-kHz range is a particularly sensitive indicator of middle-ear status, in agreement with the spectral composition of the output predictor from the multivariate analyses. When tympanometric and YR tests were combined, the resulting predictor performed slightly better or the same as the predictor calculated from the use of the YR test alone. The main conclusion is that these multivariate acoustic tests of the middle ear, which are analyzed using a clinical decision theory, are effective predictors of conductive hearing loss.  相似文献   
90.
An insert ear-canal probe including sound source and microphone can deliver a calibrated sound power level to the ear. The aural power absorbed is proportional to the product of mean-squared forward pressure, ear-canal area, and absorbance, in which the sound field is represented using forward (reverse) waves traveling toward (away from) the eardrum. Forward pressure is composed of incident pressure and its multiple internal reflections between eardrum and probe. Based on a database of measurements in normal-hearing adults from 0.22 to 8 kHz, the transfer-function level of forward relative to incident pressure is boosted below 0.7 kHz and within 4 dB above. The level of forward relative to total pressure is maximal close to 4 kHz with wide variability across ears. A spectrally flat incident-pressure level across frequency produces a nearly flat absorbed power level, in contrast to 19 dB changes in pressure level. Calibrating an ear-canal sound source based on absorbed power may be useful in audiological and research applications. Specifying the tip-to-tail level difference of the suppression tuning curve of stimulus frequency otoacoustic emissions in terms of absorbed power reveals increased cochlear gain at 8 kHz relative to the level difference measured using total pressure.  相似文献   
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