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81.
Joseph G. Major Jr. Melinda E. Wales John E. Houghton Julie A. Maley Jeffrey N. Davidson James R. Wild 《Journal of molecular evolution》1989,28(5):442-450
Summary Aspartate transcarbamoylase (ATCase, EC 2.1.3.2) is the first unique enzyme common to de novo pyrimidine biosynthesis and
is involved in a variety of structural patterns in different organisms. InEscherichia coli, ATCase is a functionally independent, oligomeric enzyme; in hamster, it is part of a trifunctional protein complex, designated
CAD, that includes the preceding and subsequent enzymes of the biosynthetic pathway (carbamoyl phosphate synthetase and dihydroorotase).
The complete complementary DNA (cDNA) nucleotide sequence of the ATCase-encoding portion of the hamster CAD gene is reported
here. A comparison of the deduced amino acid sequences of the hamster andE. coli catalytic peptides revealed an overall 44% amino acid similarity, substantial conservation of predicted secondary structure,
and complete conservation of all the amino acids implicated in the active site of theE. coli enzyme. These observations led to the construction of a functional hybrid ATCase formed by intragenic fusion based on the
known tertiary structure of the bacterial enzyme. In this fusion, the amino terminal half (the “polar domain”) of the fusion
protein was provided by a hamster ATCase cDNA subclone, and the carboxyl terminal portion (the “equatorial domain”) was derived
from a clonedpyrBI operon ofE. coli K-12. The recombinant plasmid bearing the hybrid ATCase was shown to satisfy growth requirements of transformedE. coli pyrB
− cells. The functionality of thisE. coli-hamster hybrid enzyme confirms conservation of essential structure-function relationships between evolutionarily distant
and structurally divergent ATCases. 相似文献
82.
83.
It had been previously reported that aromatic amines were not substrates for the bacterial quinoprotein methylamine dehydrogenase. In this study, benzylamine-dependent activity was also not observed in the steady-state assay of this enzyme with the artificial electron acceptor phenazine ethosulfate (PES). Benzylamines did, however, stoichiometrically reduce the protein-bound tryptophan tryptophylquinone (TTQ) prosthetic group and acted as reversible competitive inhibitors of methylamine oxidation when the enzyme was assayed with PES. When methylamine dehydrogenase activity was monitored using a steady-state assay which employed its physiological electron acceptor amicyanin instead of PES, very low but detectable benzylamine-dependent activity was observed. The reactions of a series of para-substituted benzylamines with methylamine dehydrogenase were examined. A Hammett plot of the log of Ki values for the competitive inhibition by these amines against sigma p exhibited a negative slope. Rapid kinetic measurements allowed the determination of values of k3 and Ks for the reduction of TTQ by each of these amines. A Hammett plot of log k3 versus sigma p exhibited a positive slope, which suggests that the oxidation of these amines by methylamine dehydrogenase proceeds through a carbanionic reaction intermediate. A negative slope was observed for the correlation between log Ks and sigma p. Plots of log k3 and log Ks against substituent constants which reflected either resonance or field/inductive parameters for each para substituent indicated that the magnitude of k3 was primarily influenced by field/inductive effects while Ks was primarily influenced by resonance effects. No correlation was observed between either k3 or Ks and the relative hydrophobicity of the para-substituted benzylamines or steric parameters.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
84.
Faster voltage-dependent activation of Na+ channels in growth cones versus somata of neuroblastoma N1E-115 cells.
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Kinetics of voltage-gated ionic channels fundamentally reflect the response of the channels to local electric fields. In this report cell-attached patch-clamp studies reveal that the voltage-dependent activation rate of sodium channels residing in the growth cone membrane differs from that of soma sodium channels in differentiating N1E-115 neuroblastoma cells. Because other electrophysiological properties of these channels do not differ, this finding may be a reflection of the difference in intramembrane electric field in these two regions of the cell. This represents a new mechanism for channels to attain a range of activities both within and between cells. 相似文献
85.
Caroline D. Perkins A.Michael Davidson Martin J. Day John C. Fry 《FEMS microbiology ecology》1994,15(1-2):33-44
Abstract The kinetics of conjugation, retrotransfer and mobilisation were studied at 5–15 min intervals between strains of Pseudomonas putida using plasmid pQKH6, isolated from river epilithon, and R300B. Transconjugants from the direct conjugation of pQKH6 and mobilisation of R300B by pQKH6 appeared rapidly, reaching maximum densities within 30–60 min of the start of both filter and liquid mating experiments. However, retrotransconjugants only appeared after a delay. This delay was short (approx. 45–60 min) in filter mating and much longer (2–5 h) for liquid mating experiments. Attempts at predicting the time course of retrotransconjugant development from (1) numbers of transconjugants from the conjugation and mobilisation experiments and (ii) mathematical models based on a mass action approach, both failed to reproduce the observed delay. It was concluded that retrotransfer did not proceed by either a one-step mechanism occuring early in conjugation or two separate conjugation and mobilisation steps. The clear demonstration of a delay in retrotransconjugant formation implies that a new mechanism must be sought. The likely importance of retrotransfer in the environment is discussed. 相似文献
86.
A Novel Mutation of the Fibrillin Gene Causing Ectopia Lentis 总被引:1,自引:0,他引:1
Ectopia lentis (EL), a dominantly inherited connective tissue disorder, has been genetically linked to the fibrillin gene on chromosome 15 (FBN1) in earlier studies. Here, we report the first EL mutation in the FBN1 gene confirming that EL is caused by mutations of this gene. So far, several mutations in the FBN1 gene have been reported in patients with Marfan syndrome (MFS). EL and MFS are clinically related but distinct conditions with typical manifestations in the ocular and skeletal systems, the fundamental difference between them being the absence of cardiovascular involvement in EL. We report a point mutation, cosegregating with the disease in the described family, that displays EL over four generations. The mutation changes a conserved glutamic acid residue in an EGF-like motif, which is the major structural component of the fibrillin and is repeated throughout the polypeptide. In vitro mutagenetic studies have demonstrated the necessity of an analogous glutamic acid residue for calcium binding in an EGF-like repeat of human factor IX. This provides a possible explanation for the role of this mutation in the disease pathogenesis. 相似文献
87.
Estimating regional carbon stocks and spatially covarying edaphic factors using soil maps at three scales 总被引:2,自引:1,他引:1
Most estimates of regional and global soil carbon stocks are based on extrapolations of mean soil C contents for broad categories
of soil or vegetation types. Uncertainties exist in both the estimates of mean soil C contents and the area over which each
mean should be extrapolated. Geographic information systems now permit spatially referenced estimates of soil C at finer scales
of resolution than were previously practical. We compared estimates of total soil C stocks of the state of Maine using three
methods: (1) multiplying the area of the state by published means of soil C for temperate forests and for Spodosols; (2) calculating
areas of inclusions of soil taxa in the 1:5,000,000 FAO/UNESCO Soils Map of the World and multiplying those areas by selected
mean carbon contents; and (3) calculating soil C for each soil series and map unit in the 1:250,000 State Soil Geographic
Data Base (STATSGO) and summing these estimates for the entire state. The STATSGO estimate of total soil C was between 23%
and 49% higher than the common coarse scale extrapolations, primarily because STATSGO included data on Histosols, which cover
less than 5% of the area of the state, but which constitute over one-third of the soil C. Spodosols cover about 65% of the
state, but contribute less than 39% of the soil C. Estimates of total soil C in Maine based on the FAO map agreed within 8%
of the STATSGO estimate for one possible matching of FAO soil taxa with data on soil C, but another plausible matching overestimated
soil C stocks. We also compared estimates from the 1:250,000 STATSGO database and from the 1:20,000 Soil Survey Geographic
Data Base (SSURGO) for a 7.5 minute quadrangle within the state. SSURGO indicated 13% less total soil C than did STATSGO,
largely because the attribute data on depths of soil horizons in SSURGO are more specific for this locality. Despite localized
differences, the STATSGO database offers promise of scaling up county soil survey data to regional scales because it includes
attribute data and estimates of areal coverage of C-rich inclusions within map units. The spatially referenced data also permit
examination of covariation of soil C stocks with soil properties thought to affect stabilization of soil C. Clay content was
a poor predictor of soil C in Maine, but drainage class covaried significantly with soil C across the state. 相似文献
88.
Emile CL. Marnette Harm Houweling Herman Van Dam Jan Willem Erisman 《Biogeochemistry》1993,23(2):119-144
The chemical composition of surface waters of two Dutch moorland pools and of incident precipitation, was monitored from 1982
to 1990. For this period, sulfur and water budgets were calculated using a hydrochemical model developed for well-mixed non-stratifying
lakes. Total atmospheric deposition of S decreased significantly after 1986 at both locations. A model describing the sulfur
budget in terms of input, output and reduction/oxidation processes predicted a fast decrease of pool water SO4
2− concentrations after a decrease of atmospheric input. However, SO4
2− concentrations in the surface water was lowered only slightly or remained constant. Apparently a source within the lake caused
the unexpectedly high SO4
2− concentrations. The possible supply of SO4
2− from the sediment through regulation by (K-)Al-SO4 containing minerals or desorption of SO4
2− from positively charged surfaces in the sediment was evaluated. Solubility calculations of pore water with respect to alunite,
basaluminite and jurbanite indicated that SO4
2− concentration was not regulated by these minerals. It is suggested here (1) that desorption of SO4
2− from peaty sediments may account for the estimated SO4
2− supply provided that the adsorption complex is periodically recharged by partial oxidation of the upper bottom sediments
and (2) that because of exposure of a part of the pool bottom to the atmosphere during dry summers and subsequent oxidation
of reduced S, the amount of SO4
2− may be provided which complements the decreasing depositional SO4
2− input. In future research these two mechanisms need to be investigated. 相似文献
89.
Submaximal,aerobic exercise training exacerbates the cardiomyopathy of postweanling Cu-depleted rats
Jeannette Davidson Denis M. Medeiros Robert L. Hamlin James E. Jenkins 《Biological trace element research》1993,38(3):251-272
To determine the dual effect of exercise training and copper depletion on myocardial function and ultrastructure, postweanling
rats were either trained or sedentary while fed copper-adequate or copper-deficient diets for 8 wk. Rats developed characteristic
myocardial subcellular degeneration and increased cardiac mitochondrial volume density when copper depleted, despite lack
of overt cardiac hypertrophy, hypertension, or anemia. Training combined with copper depletion induced mild left ventricular
hypertrophy. Basal laminae appeared fractionated in areas at capillary-myocyte interface, with focal pericapillary and interstitial
collagen accumulation, where-as overt fibrosis was absent or minimal. Electrocardiograms revealed increased QRS wave and QT
duration and notching of QRS complex with copper depletion, consistent with intraventricular conductance disturbances. The
oxidative capacity of soleus muscle increased with training in copper-adequate rats, but was reduced with progressive copper
depletion. These data suggest that copper depletion and training are synergistic in effecting focal accumulation of collagen,
with deleterious effect on exercise capacity. 相似文献
90.