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101.
An experimental approach and methodology are described for determining the reactive properties and ionization constants of individual functional groups of proteins within biomolecular complexes. The ionization constants and reactivities of the methionyl-l amino terminus and the lysyl-3 residue of the alanine rich 50S ribosomal "A" protein from Bacillus stearothermophilus have been determined by an extension of the competitive labeling technique used by H. Kaplan, K. J. Stevenson, and B. S. Hartley ((1971), Biochem. J. 124, 289-299). This approach employs (1-14C)- and (3H)acetic anhydride in a double-labeling procedure. In 0.1 M KCl-0.02 M Mg2+-0.05 M Veronal at 10 degrees the methionyl-l amino terminus has a pKa of 7.5 and is exposed on the surface of the ribosome. The lysyl-3 has a pKa of 10 and is also exposed to solvent at the surface of the 50S subunit. Based on a linear free energy relationship (Bronsted plot) obtained with a series of standard amines the methionyl amino terminus has a substantially higher reactivity than expected from its ionization constant. The lysyl epsilon-amino group has the expected reactivity. The abnormally high reactivity of the methionyl amino terminus can only be accounted for by a specific interaction with other functional groups in the ribosome. These data support the proposal that the charged state of this residue is important in the structure and function of the "A" protein at the surface of the ribosome. 相似文献
102.
A lambda lysogen with the prophage inserted into the arabinose B gene of Escherichia coli strain K-12 has been prepared. Induction of the phage from this lysogen yields viable phage at a frequency 4 X 10(-6) that found for induction of lysogens with phage inserted at the normal attachment site. Over 30% of the phage particles induced from the insertion in ara are arabinose-transducing phage. The excision end points of 62 independently isolated, nondefective araC-transducing phage containing less than the entire araC gene were genetically determined and were found to be randomly distributed through the araC gene. The amount of arabinose deoxyribonucleic acid contained on four selected transducing phage was determined by electron microscopy of deoxyribonucleic acid heteroduplexes, providing a physical map of the araC gene. The efficiency with which these phage transduce araC and araB point mutations was found to be approximately proportional to the homology length available for recombination. 相似文献
103.
Nicotinamide adenine dinucleotide-linked malate dehydrogenase has been purified from Pseudomonas testosteroni (ATCC 11996). The purification represents over 450-fold increase in specific activity. The amino acid composition of the enzyme was determined and found to be quite different from the composition of the malate dehydrogenases from animal sources as well as from Escherichia coli. Despite this difference, however, the data show that the enzymatic properties of the purified enzyme are remarkably similar to those of other malate dehydrogenases that have been previously studied. The Pseudomonas enzyme has a molecular weight of 74,000 and consists of two subunits of identical size. In addition to L-malate, the enzyme slowly oxidizes other four-carbon dicarboylates having an alpha-hydroxyl group of S configuration such as meso- and (-) tartrate. Rate-determining steps, which differ from that of the reaction involving L-malate, are discussed for the reaction involving these alternative substrates. Oxidation of hydroxymalonate, a process previously undetected with other malate dehydrogenases, is demonstrated fluorometrically. Hydroxymalonate and D-malate strongly enhance the fluorescence of the reduced nicotinamide adenine dinucleotide bound to the enzyme. The enzyme is A-stereospecific with respect to the coenzyme. Malate dehydrogenase is present in a single form in the Pseudomonas. The susceptibility of the enzyme to activation or inhibition by its substrates-particularly the favoring of the oxidation of malate at elevated concentrations-strongly resembles the properties of the mitochondrial enzymes. The present study reveals that whereas profound variations in chemical composition have occurred between the prokaryotic and eukaryotic enzymes, the physical and catalytic properties of malate dehydrogenase, unlike lactate dehydrogenase, are well conserved during the evolutionary process. 相似文献
104.
105.
Continuous exposure to dichlorvos (DDVP) vapors from birth in rats caused a statistically significant mean delay of 10 da in the onset of the first estrous cycle. DDVP, BUT NOT ITS PRIMARY METABOLITE (DES-METHYL DDVP), could be readily isolated from blood and ovarian tissue, and occassionally in minute quantities from the kidney and adrenal tissues. DDVP was recovered only once in 40 brain tissue samples examined. 相似文献
106.
107.
Some physiological characteristics of a mutant (E1) of Anacystis nidulans R2, incapable of growing at air level of CO2, are described. E1 is capable of accumulating inorganic carbon (Ci) internally as efficiently as the wild type (R2). The apparent photosynthetic affinity for Ci in E1, however, is some 1000 times lower than that of R2. The kinetic parameters of ribulose 1,5-bisphosphate carboxylase/oxygenase from E1 are similar to those observed in R2. The mutant appears to be defective in its ability to utilize the intracellular Ci pool for photosynthesis and depends on extracellular supply of Ci in the form of CO2. The very high apparent photosynthetic Km (CO2) of the mutant indicate a large diffusion resistance for CO2. Data obtained here are used to calculate the permeability coefficient for CO2 between the bulk medium and the carboxylation site of cyanobacteria. 相似文献
108.
109.
Jonathan Kaplan 《Biology & philosophy》2008,23(5):625-638
The concepts of adaptive/fitness landscapes and adaptive peaks are a central part of much of contemporary evolutionary biology;
the concepts are introduced in introductory texts, developed in more detail in graduate-level treatments, and are used extensively
in papers published in the major journals in the field. The appeal of visualizing the process of evolution in terms of the
movement of populations on such landscapes is very strong; as one becomes familiar with the metaphor, one often develops the
feeling that it is possible to gain deep insights into evolution by thinking about the movement of populations on landscapes
consisting of adaptive valleys and peaks. But, since Wright first introduced the metaphor in 1932, the metaphor has been the
subject of persistent confusion, from equivocation over just what the features of the landscape are meant to represent to
how we ought to expect the landscapes to look. Recent advances—conceptual, empirical, and computational—have pointed towards
the inadequacy and indeed incoherence of the landscapes as usually pictured. I argue that attempts to reform the metaphor
are misguided; it is time to give up the pictorial metaphor of the landscape entirely and rely instead on the results of formal
modeling, however difficult such results are to understand in ‘intuitive’ terms.
相似文献
Jonathan KaplanEmail: |
110.
The liver peptide hepcidin regulates body iron, is upregulated in iron overload and inflammation, and is downregulated in iron deficiency/hypoxia. The transmembrane serine protease matriptase-2 (TMPRSS6) inhibits the hepcidin response and its mutational inactivation causes iron-deficient anemia in mice and humans. Here we confirm the inhibitory effect of matriptase-2 on hepcidin promoter; we show that matriptase-2 lacking the serine protease domain, identified in the anemic Mask mouse (matriptase-2(MASK)), is fully inactive and that mutant R774C found in patients with genetic iron deficiency has decreased inhibitory activity. Matriptase-2 cleaves hemojuvelin (HJV), a regulator of hepcidin, on plasma membrane; matriptase-2(MASK) shows no cleavage activity and the human mutant only partial cleavage capacity. Matriptase-2 interacts with HJV through the ectodomain since the interaction is conserved in matriptase-2(MASK). The expression of matriptase-2 mutants in zebrafish results in anemia, confirming the matriptase-2 role in iron metabolism and its interaction with HJV. 相似文献