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51.
A method is introduced to identify amino acid residues that dictate the functional diversity acquired during evolution in a protein family. Using over 80 enzymes of the chymotrypsin family, we demonstrate that the general organization of the phylogenetic tree and its functional branch points are fully accounted for by a limited number of residues that cluster around the active site of the protein and define the contact region with the P1-P4 residues of substrate. 相似文献
52.
Histamine formation in rat brain during development 总被引:2,自引:7,他引:2
J C Schwartz C Lampart C Rose M C Rehault S Bischoff H Pollard 《Journal of neurochemistry》1971,18(9):1787-1789
53.
K Rose M G Simona L A Savoy P O Regamey B N Green G M Clore A M Gronenborn P T Wingfield 《The Journal of biological chemistry》1992,267(27):19101-19106
Ner protein of bacteriophage Mu, produced by recombinant DNA techniques in Escherichia coli, has been found to possess a molecule of pyruvic acid attached covalently through carbon-2 to the amino-terminal cysteine residue. The intact protein and the amino-terminal chymotryptic peptide were found by mass spectrometry to be 70 mass units heavier than expected. The modified peptide was unstable under mildly acid or mildly basic conditions. Two-dimensional nuclear magnetic resonance spectroscopy of the modified and unmodified forms of the amino-terminal chymotryptic peptide was consistent with the presence of pyruvate linked through carbon-2 to the amino-terminal Cys residue. Treatment of the modified form with 2,4-dinitrophenylhydrazine in acid medium led to the expected hydrazone of pyruvic acid, which was identified by high pressure liquid chromatography. Of the two proteins known to be modified by pyruvate through its central carbon (the other being human adult hemoglobin, in which the modified form represents only a very minor fraction), Ner is the first protein found to be modified quantitatively. Given the instability of the modification, it may be more prevalent than recognized hitherto. Incubation with 2,4-dinitrophenylhydrazine may offer a useful means of detecting the presence of pyruvate linked to proteins in this way. 相似文献
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Gary Parkinson Simon Gaisford Qian Ru Alastair Lockwood Ashkan Khalili Rose Sheridan Peng T. Khaw Steve Brocchini Hala M. Fadda 《AAPS PharmSciTech》2012,13(4):1063-1072
We are developing tablet dosage forms for implantation directly into the subconjunctival space of the eye. The matrix metalloproteinase inhibitor, ilomastat, has previously been shown to be efficacious at suppressing scarring following glaucoma filtration surgery (GFS). We report on the physical characterisation of ilomastat which is being developed for ocular implantation. Since ilomastat is being considered for implantation it is necessary to examine its polymorphs and their influence on aspects of the in vitro drug release profile. X-ray powder diffraction identified two polymorphs of ilomastat from different commercial batches of the compound. Tablets were prepared from the two different polymorphs. Isothermal perfusion calorimetry was used to show that amorphous content is not increased during tablet formulation. The melting points of the two polymorphs are 188 and 208°C as determined by differential scanning calorimetry. Utilising single crystal X-ray diffraction, the structural conformations and packing arrangements of the different polymorphs were determined. The orthorhombic crystal crystallised as a monohydrate while the second monoclinic crystal form is non-solvated. Ilomastat tablets prepared from the two different solid forms exhibited similar drug release profiles in vitro under conditions mimicking the aqueous composition, volume and flow of the subconjunctival space after GFS. This suggests that a reproducible dose at each time point during release after implantation should be achievable in vivo with ilomastat tablets prepared from the two polymorphs identified. 相似文献
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College undergraduates classified as high (n = 25) and low (n = 25) on recent life stress participated in an experiment involving a novel laboratory stressor. Heart rate and pulse arrival time (PAT) were measured during baseline, anticipation, testing, and recovery periods of the experiment. The results did not replicate those obtained by Pardine and Napoli in that high and low life stress subjects did not show differential physiological reactions. In addition, regression analyses failed to demonstrate that physiological reactivity moderated the relationship between life stress and subsequent self-reported psychiatric or physical health symptomatology. The present findings demonstrated neither the stress-buffering effects of physiological reactivity nor a relationship between life stress and reactivity when the latter was conceptualized as an outcome. 相似文献
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The evolutionary relationships between three major components of Darwinian fitness, development rate, growth rate and preadult
survival, were estimated using a comparison of 55 distinct populations ofDrosophila melanogaster variously selected for age-specific fertility, environmental-stress tolerance and accelerated development. Development rate
displayed a strong net negative evolutionary correlation with weight at eclosion across all selection treatments, consistent
with the existence of a size-versus-time tradeoff between these characters. However, within the data set, the magnitude of
the evolutionary correlation depended upon the particular selection treatments contrasted. A previously proposed tradeoff
between preadult viability and growth rate was apparent only under weak selection for juvenile fitness components. Direct
selection for rapid development led to sharp reductions in both growth rates and viability. These data add to the mounting
results from experimental evolution that illustrate the sensitivity of evolutionary correlations to (i) genotype-by-environment
(G X E) interaction, (ii) complex functional-trait interactions, and (iii) character definition. Instability, disappearance
and reversal of patterns of genetic covariation often occur over short evolutionary time frames and as the direct product
of selection, rather than some stochastic process. We suggest that the functional architecture of fitness is a rapidly evolving
matrix with reticulate properties, a matrix that we understand only poorly. 相似文献