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H Decker

Publications and source records attributed to H Decker.

70 records · Page 4Linked to original sources
PubMed1985

Binding of oxygen and carbon monoxide to arthropod hemocyanin: an allosteric analysis.

The binding of oxygen and carbon monoxide to hemocyanin from the mangrove crab Scylla serrata and the lobster Homarus americanus has been studied by thin-layer optical absorption and front face fluorescence techniques. Three types of experiments were performed on subunit and oligomeric preparations of each hemocyanin: oxygen binding, carbon monoxide binding, and oxygen-carbon monoxide competition studies. The results obtained from the subunit preparations of dissociated oligomers from both hemocyanins show that the binding site can be ligated by either one oxygen or one carbon monoxide. The binding results obtained with the oligomeric samples of hemocyanin from both species cannot be described by the two-state MWC model [Monod, J., Wyman, J., & Changeux, J. P. (1965) J. Mol. Biol. 12, 88-118] since the data from the three types of binding experiments cannot be fit with a single set of binding constants. The MWC model has been extended by including a third allosteric form, and an analysis based on the three-state model is able to fit the data from the three types of experiments with the same set of binding constants. The comparison of the oxygen to carbon monoxide affinity ratios (kO2/kCO) indicates that the structure around the binding site of subunits in the T form oligomer is similar to that of the free subunits. The oligomeric forms of both these hemocyanins bind carbon monoxide with a weak but definite positive cooperativity. An analysis of the affinity ratios for the T, S, and R forms suggests that the high affinity of the R form results from a specific interaction between oxygen and binding site.

Allosteric Regulation↗
PubMed1985

Classification of sulpiride, clozapine and haloperidol by toposelective recording from different brain structures in the immobilized rat (stereo-EEG).

The formerly designed method for quantitative evaluation of drug effects in rats is employed for the classification of three different neuroleptic drugs, namely sulpiride, clozapine and haloperidol. Analysis of the power density spectra as obtained from the recorded activity in the cortex, striatum and reticular core results in a 'fingerprint' for each drug, which is then compared to the known data on various standard drugs by means of a stepwise discriminant analysis. After cumulative dosage dose-response curves are given for single frequency bands and different brain areas. The ED50 values are close to the drug dosages applied to humans. All three neuroleptics can be differentiated from each other, but are classified very close to low dosages of chlorpromazine according to the classification rule established in an earlier paper. Relying on the overall difference of the drug's action in comparison to placebo, a single dose-response relationship is given for each compound.

Animals↗
PubMed1984

Classification of drugs by stereotactic recording of focal brain activity in the rat (stereo-EEG).

A new electrophysiological model is described in which different types of psychoactive drugs can be differentiated from each other in the rat. Simultaneous focal recording of electrical activity from three brain regions in the presence of accumulating drug actions is followed by a calculation of the EEG power density spectrum. Division of the spectrum into 5 frequency bands for each brain region allows to evaluate a total of 15 variables. The values obtained after the last of 8 drug dosages are taken for a stepwise discriminant analysis. A distinct separation of amphetamine, chlorpromazine, diazepam and imipramine from placebo is achieved. The discriminant function obtained will serve for further drug classification.

Amphetamine↗
PubMed1983

Positive cooperativity in binding carbon monoxide to hemocyanin.

The binding of carbon monoxide to hemocyanin from the crab Scylla serrata has been studied by thin layer optical absorption and front face fluorescence techniques. The binding to the monomeric form is completely noncooperative whereas the binding to the native oligomeric form is found to be weakly but definitely cooperative. An analysis based on the MWC model of the oxygen and carbon monoxide binding curves indicates that the allosteric constant, L, describing the equilibrium between the 2 unligated forms is different for each ligand. This implies that at least 3 allosteric forms are needed to characterize the binding of oxygen and carbon monoxide to this hemocyanin.

Allosteric Regulation↗
PubMed1983

Thin-layer fluorescence cell for ligand binding studies.

A high precision method for measuring the binding of gaseous ligands to proteins is presented. Front face fluorescence techniques are utilized with a special thin-layer cell in order to monitor the change in fluorescence intensity caused by changing the ligand partial pressure. The method is illustrated by examining the binding of carbon monoxide to hemocyanin from the lobster Homarus americanus.

Animals↗
PubMed1982

Hemocyanins in spiders, XV. The role of the individual subunits in the assembly of Eurypelma hemocyanin.

The role of the seven different subunits in the quaternary structure of the 24-meric (37 S) hemocyanin of the tarantula, Eurypelma californicum, was studied by reassembly experiments. Individual subunits and combinations of 2, 3, 4, etc. different subunits were incubated in a total concentration of 1-2 mg/ml overnight in Tris buffer, pH 7.5, omitting divalent cations. The reassembly mixtures were then analyzed by thinlayer gel filtration, electron microscopy, analytical ultracentrifugation, and polyacrylamide gel electrophoresis. At least at the low protein concentration employed, none of the isolated monomeric subunits (a, d, e, f, g) is capable of hexamer are likewise ineffective. The minimal number of monomers required for hexamer formation is three, but only about half of the 10 possible combinations was effective. The best results were obtained with a + f + g. Four and five monomers always yielded hexamers, although the composition of the latter is not known. The heterodimer bc is a prerequisite to go beyond the hexameric state. bc alone forms tetramers and large, strand-like aggregates. a shows a high affinity towards bc, trimers and tetramers being formed. Out of all combinations of bc and two monomers, only one (bc + a + g) produced hexamers or heptamers. With three monomers plus bc, dodecamers and larger structures (but no 24-mers) were obtained, but only if also a was present (bc + a + f + g being the best combination). If all subunits minus one monomer were combined, the hexamer/heptamer level was not exceeded if a was omitted. 24-mers were formed in appreciable yield only in one case, namely if e was absent. However, this reassembly product turned out to be unstable. To obtain stable 24-meric hemocyanin, the complete set of subunits had to be present. The roles of the different subunits are defined as follows: bc is a "core" subunit which forms the inter-hexamer link within each dodecameric half. It serves also in inter-dodecamer bonding. The artificial homodimers bb and cc are ineffective in this respect. bc must be supplemented by a which is needed for dodecamer stabilization. f is also required to link two dodecamers, while g serves to stabilize this bridge. d and e are required to finish off and to stabilize the 24-mer. A model is proposed showing some neighbourhood relationships of the subunits within the hemocyanin 24-mer, and a pathway of reassembly is discussed.

Animals↗
PubMed1981

On the role of dimeric subunits in the quaternary structure of arthropod hemocyanins.

Partial alkaline dissociation of 24 S (12-meric), 35 S (24-meric) and 60 S (48-meric) hemocyanin from various arthropods was studied by polyacrylamide gradient gel electrophoresis and, in some cases, by electron microscopy. If there are no stable dimers among the subunits, dissociation starts by cleavage of interhexamer bonds, leading to intermediates which are hexamers or multiples of hexamers. Whenever a hemocyanin contained stable dimers, inter-hexamer bonds were also very stable as indicated by the formation of 30 S (19-meric) or 18 S (7-eric) intermediates as primary products. In such cases, inter-hexamer bonds could be cleaved by treatment with reducing agents or with 4M urea; correspondingly, these agents also cleaved the respective dimers into the constituent polypeptide chains. It is concluded that in all cases the dimeric subunits function as bridges between hexamers.

Animals↗
PubMed1980

Hemocyanins in spiders, IX. Homogeneity, subunit composition and the basic oligomeric structure of Eurypelma californicum hemocyanin.

37S hemocyanin isolated from the tarantula Eurypelma californicum was subjected to a variety of fractionation procedures: Electrophoresis in polyacrylamide gels, isoelectrofocusing, and ion exchange chromatography. Single fractions were dissociated at alkaline pH and/or by sodium dodecyl sulfate and the resulting subunits separated by polyacrylamide gel electrophoresis in gradient slab gels followed, in many cases, by quantitative estimation of the subunit bands. In addition, crossed immunoelectrophoresis was employed to analyze the subunit composition. Separation of the native hemocyanin into fractions with different subunit composition was not observed. Also, hemocyanin samples taken from individual spiders showed a strikingly uniform composition. It is concluded that Eurypelma hemocyanin is homogeneous for all practical means. This implies that the seven different polypeptide chains described previously for Eurypelma hemocyanin all take part in the formation of the 24 subunits, 37S oligomeric molecule. By monitoring chromatographic effluents at 280 nm, scanning stained electrophoresis gels and determining peak areas in these experiments and in crossed immunoelectrophoresis, the relative quantities of the seven different polypeptide chains in the whole molecule were determined as follows (number per 37S molecule): 6a, 2b, 2c2, 2c4, 4d, 4e, 4f. The ratio between a and c2 (6:2) is the least well established. Since 7 different polypeptide chains are incorporated, the hexameric molecule cannot represent the basic oligomeric structure in tarantula hemocyanin, but probably the dodekamer. If Eurypelma hemocyanin is incubated with sucrose, partial dissociation occurs, yielding a fragment sedimenting with ca. 28S. This process is reversible. The 28S fragment still contains all subunit species; it is thought to represent a (3 x 6) or closely related structure. The implications of this finding with respect to the symmetry of the native hemocyanin molecule are discussed.

Animals↗
PubMed1980

Hemocyanins in spiders, XII. Dissociation and reassociation of Eurypelma hemocyanin.

The dissociation and reassembly of Eurypelma californicum hemocyanin was studied under various conditions of pH, ionic strength and protein, calcium and magnesium concentrations. The hemolymph concentrations of calcium and magnesium were determined to be ca. 4 and 0.9mM, respectively. Eurypelma hemocyanin does not dissociate upon dilution down to 0.04 mg/ml. At physiological pH, phosphate causes partial dissociation. Dissociation at alkaline pH requires two days for completion and is dependent on hemocyanin concentration. Reassociation, starting with the total dissociated subunits, yielded 37 S, 16 S and unreacted 5 S material. The yield of 37 S particles after one day was 75%, but rose slowly to 84% after two weeks. Raising the calcium concentration above 1mM resulted in an increase of 16 S particles (hexamers). High magnesium concentrations interfered with ordered reassembly. However, 37 S, native-like particles are readily formed in the absence of both calcium and magnesium. Reassociation is strongly favoured if ionic strength is increased. The significance of the 16 S reassociation product was studied by repeated dissociation and reassociation and analysis of subunit composition. It was found that the 37 S reassociate contained the whole set of subunits, their quantitative proportions being identical compared to native 37 S hemocyanin. In contrast, the 16 S material was lacking the dimeric subunit bc4 (= 4D). Evidently, this hexameric product arises by incorrect reassociation of monomeric subunits. P50 and nH values were recorded for the reassociated hemocyanins. The 37 S component had a higher affinity and lower cooperativity than native hemocyanin, possibly due to ageing of the subunits.

Animals↗
PubMed1980

[Some observations on the influence of ecological factors on the occurrence and the distribution of plant parasitic nematodes in the Democratic Republic of Sudan].

Among the various ecological factors influencing the phytonematode fauna the types of plant and soil determine the nematode range. Radopbolus similis, Helicotylencbus multicinctus, Hemicriconemoides strictatbecatus, Ditylencbus augustus, Tylencbulus semipenetrans, and possibly Paratropburus kenanae are examples of high host specificity. The Pratylencbus, Meloidogyne, Xipabenema, as well as certain Tylencborbyncbus species differ in their demands to the soils. Non-irrigated natural locations exhibit a direct dependence of the range of species on the amount of precipitation, as can be seen from examples from the < 300 mm, 300 to 400 mm, and > 500 mm- annual precipitation ranges. Regions irrigated regularly correspond to the last-mentioned group. The moisture content depending on the annual floodings in the Gash and Baraka deltas has to be classified as a separate and special case. There are no sufficient findings available on the influence of the temperature and of the harmful organisms. They exert a stronger influence on the population density than on the range of species.

Animals↗
PubMedDate not supplied

Genetic control of polyketide biosynthesis in the genus Streptomyces.

The genetic control of polyketide metabolite biosynthesis in Streptomyces sp. producing actinorhodin, daunorubicin, erythromycin, spiramycin, tetracenomycin and tylosin is reviewed. Several examples of positively-acting transcriptional regulators of polyketide metabolism are known, including some two-component sensor kinase-response regulator systems. Translational and posttranslational control mechanisms are only briefly mentioned since very little is known about either of these processes. Examples of how enzyme levels and substrate supply affect polyketide metabolism also are discussed.

Amino Acid Sequence↗
PubMedDate not supplied

Comparison of relative renal function measured with either 99mTc-DTPA or 99mTc-EC dynamic scintigraphies with that measured with 99mTc-DMSA static scintigraphy.

OBJECTIVE: The aim of this study was to compare the renal function measured with either 99mTc-DTPA or 99mTc-EC dynamic scintigraphies with that measured using 99mTc-DMSA static scintigraphy. METHODS: the values of relative renal function measured in 111 renal dynamic scintigraphies performed either with 99mTc-DTPA (55 studies) or with 99mTc-EC (56 studies) were compared with the relative function measured using 99mTc-DMSA static scintigraphy performed within a 1-month period. The comparisons were performed using Wilcoxon signed rank test. The number of 99mTc-DTPA and 99mTc-EC studies that presented relative renal function different by more than 5% from that measured with 99mTc-DMSA, using chi square test were also compared. RESULTS: the relative renal function measured with 99mTc-EC is not statistically different from that measured with 99mTc-DMSA (p = 0.97). The relative renal function measured with 99mTc-DTPA was statistically different from that measured using 99mTc-DMSA, but with a borderline statistical significance (p = 0.05). The number of studies with relative renal function different by more than 5% from that measured with 99mTc-DMSA is higher for the 99mTc-DTPA scintigraphy (p = 0.04) than for 99mTc-EC. CONCLUSION: the relative renal function measured with 99mTc-EC dynamic scintigraphy is comparable with that measured with 99mTc-DMSA static scintigraphy, while the relative renal function measured with 99mTc-DTPA dynamic scintigraphy presents a significant statistical difference from that measured with 99mTc-DMSA static scintigraphy.

Adolescent↗