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Biomedical subjects

R Santucci

Publications and source records attributed to R Santucci.

At least 91 records · Page 5Linked to original sources
PubMed1989

Spatial attentional shifts: further evidence for the role of polysensory mechanisms using visual and tactile stimuli.

Normal subjects performed simple reaction time responses to lateralized visual target stimuli (Experiment 1) and lateralized tactile target stimuli (Experiment 2). In each experiment, the lateralized targets were preceded at one of four intervals by a visual or tactile cue located on the same (valid cue), or opposite (invalid cue) side, or on both sides (neutral cue). The validity of the visual and tactile cues influenced the speed of response to either target stimulus. These findings, together with those previously reported (Buchtel and Butter, Neuropsychologia 26, 499-509, 1988), are consistent with the view that intra- and inter-modal spatial cueing is effective with modalities that are linked to orienting systems in which movements of the sensory array serve to improve sensory analysis.

Adult↗
PubMed1988

Multiplicity of interactions between dromedary hemoglobin and solvent components. A structural and functional study.

The interaction of dromedary hemoglobin with various solvent components [2-(p-chlorophenoxy)-2-methylpropionic acid (CFA), 2,3-bisphospho-D-glycerate (glycerate-2,3-P2) and chloride] has been studied. 1. CFA greatly lowers the oxygen affinity of dromedary hemoglobin. 2. The oxygen-linked CFA binding sites are probably located in the deoxy derivative at the alpha cleft, while in the oxy form and in the presence of two other effectors (glycerate-2,3-P2 and chloride) additional, structurally and possibly functionally relevant binding site(s) should be considered. 3. Both CFA and glycerate-2,3-P2 stabilize the deoxy-like tertiary structure in the oxy derivative. 4. Chloride appears to be fundamental to obtain quaternary structural changes. 5. Interaction energy, retained in the protein when the three ligands (CFA, glycerate-2,3-P2 and chloride) are bound to the oxy form, favours intermediates not stable if only one or two allosteric effector(s) is (are) present on the protein. 6. The oxygen affinity appears to be related to both tertiary and quaternary structural changes, while cooperatively is largely invariant with solvent conditions. In conclusion, the functional properties of dromedary hemoglobin do not depend in any simple way on the variety of stabilized conformations.

2,3-Diphosphoglycerate↗
PubMed1988

CD studies on the reversed heme orientation in monomeric Glycera dibranchiata hemoglobins.

Circular dichroism spectra of three monomeric components of Glycera dibranchiata hemoglobins are reported. Contrary to what is found for most hemoglobins and myoglobins, G. dibranchiata hemoglobins display largely negative dichroic spectra in the Soret region. Independent NMR measurements have shown that the same monomeric hemoglobin components contain the heme moiety predominantly (greater than 85%) oriented in a reversed way with respect to the orientation which occurs in most hemoglobins and myoglobins. On the basis of these independent NMR studies, and also of previous data on other invertebrate hemoproteins, a correlation appears evident between reversed heme orientation in hemoglobins and negative ellipticity in the Soret CD spectrum. This represents a simple tool to evaluate this aspect of heme asymmetric environment.

Animals↗
PubMed1988

Effect of polyanions on the spectroscopic properties of the nitric oxide derivative of ferrous dromedary (Camelus dromedarius) hemoglobin.

The effect of inositol hexakisphosphate, 2,3-diphosphoglycerate, dextran sulphate, and heparin on the spectroscopic (absorbance, circular dichroism, EPR) properties of the nitric oxide derivative of ferrous dromedary (Camelus dromedarius) hemoglobin was investigated. The results obtained show that: (i) all polyanions bind to the protein at the same sites, but with different affinities; (ii) polyanions affect the protein conformation of the ferrous nitrosyl derivative in a different way with respect to aquo-ferric and ferrous oxy dromedary hemoglobin; and (iii) the data obtained provide further independent evidence for the existence in dromedary hemoglobin of two functionally distinct polyanion binding sites that affect the conformational equilibrium of the protein in opposite ways.

2,3-Diphosphoglycerate↗
PubMed1988

Multiplicity of different stable quaternary conformations in ferric hemoglobin from Camelus dromedarius. Modulation by small anions.

The effect of small anions in modulating polyphosphate-induced conformational changes in ferric dromedary hemoglobin (Camelus dromedarius) was investigated. The results obtained indicate that: i) in solution, the simultaneous presence of polyphosphate and small anions is essential to obtain quaternary changes in the protein; ii) the charge, rather than the size or sterical configuration, mainly affects the quaternary structural equilibrium of the macromolecule; iii) three distinct, stable quaternary conformers of ferric dromedary hemoglobin were detected spectroscopically as a function of charge and concentration of small anion.

Animals↗
PubMed1987

Redox properties of components I and IV of trout hemoglobins: kinetic and potentiometric studies.

Redox properties of component I and IV from trout hemoglobin (Salmo irideus) have been studied kinetically and at equilibrium. In the case of component I of trout hemoglobin, the mid-point potential (Eh) is pH independent below the acid-alkaline transition (pKa approximately equal to 8.6) and decreases at higher pH, following the deprotonation of the water molecule. Similarly to human hemoglobin, the mid-point potential of component IV of trout hemoglobin is pH-dependent, but the redox Bohr effect is extended to more acid pH. Moreover, the cooperativity of the redox equilibrium process is higher than in human hemoglobin. These features parallel the oxygen-binding properties of the same hemoglobin components from trout hemolysate. Differently from human hemoglobin, the oxidation kinetics of the two hemoglobins from trout by potassium ferricyanide show markedly biphasic progress curves with pH-independent second-order rate constants. This behavior suggests a different energy barrier for the interaction with ferricyanide in the two types of subunit of both Hb components from trout.

Animals↗
PubMed1987

Unfolding and flexibility in hemoproteins shown in the case of carboxymethylated cytochrome c.

A circular dichroism study of carboxymethylated cytochrome c has been performed to obtain further information on the structural basis responsible for the observed changes in ligand binding and redox properties of the modified cytochrome c. The results give additional evidence of local structural changes occurring in the heme environment upon rupture of the (Met-80)-heme iron bond in the modified protein. This produces no alterations of the overall molecular conformation, but results in drastic changes in redox potential. In addition, analysis of the reversible conformational transitions induced by urea in the native and the modified proteins supports the idea that the modified derivative can be considered as an 'intermediate state' between the native and the fully unfolded protein.

Animals↗
PubMed1987

Structural and functional effects of selective chemical modifications of Scapharca inaequivalvis haemoglobins in relation to their unique assembly.

The structural and functional roles of lysyl and thiol groups in the dimeric (HbI) and tetrameric (HbII) haemoglobins from the mollusc Scapharca inaequivalvis have been assessed. In these haemoglobins a unique mode of assembly (the haem-carrying E and F helices form the intersubunit contact of the dimeric unit) is associated with co-operative oxygen binding. Extensive acylation is accompanied by significant haem oxidation. Modification of one or two lysyl residues per chain (corresponding to approximately 20% of the total residues) does not affect the structural and functional properties of both haemoglobins, in line with the proposal that the intersubunit contacts are rich in hydrophobic residues. The modification of the thiol groups does not influence the state of association in both HbI and HbII, despite the location of the cysteine residue common to all polypeptide chains in the vicinity of the major intersubunit contact. The effect on the functional properties depends on the size of the thiol reagent: p-chloromercuribenzoate and phenylmercuric acetate increase the oxygen affinity about 20-fold, but iodoacetamide and mercuric chloride have no effect. Moreover, electrophoresis experiments indicate that p-chloromercuribenzoate is bound in a co-operative fashion, the degree of co-operativity being much higher in the dimeric HbI. Thus, only in HbII are intermediates containing substoichiometric amounts of p-chloromercuribenzoate formed in significant amounts. Their oxygen binding properties show that reaction of only one thiol group/tetramer suffices to alter the oxygen affinity of the molecule.

Acylation↗
PubMed1986

Conformational changes induced by polyanions in haemoglobin from the dromedary (Camelus dromedarius). Role of chloride ions.

The role of chloride ions in modulating polyanion-induced conformational changes in haemoglobin from the dromedary (Camelus dromedarius) has been investigated. The results obtained have shown that: in the ferric derivative at pH 6.5 the effect of single polyanion (dextran sulphate and inositol hexakisphosphate) on the conformation is essentially local, thus involving only the tertiary structure of the protein; the presence of chloride ions at a concentration close to the physiological value (i.e. 150 mM) is essential to induce quaternary conformational changes in the polyanion-ferric protein system; comparison between structural and functional data correlates polyanion-induced tertiary conformational changes with changes in the value of midpoint potential, E'0, and quaternary changes with co-operativity.

Animals↗
PubMed1986

Conformational changes induced by polyanions in haemoglobin from Camelus dromedarius. Studies on the ferric derivatives.

The effect of inositol hexakisphosphate on the redox equilibria and on the c.d. spectra of ferric derivatives of haemoglobin from Camelus dromedarius has shown that: two distinct functionally relevant binding sites for polyanions are present on the protein; conformational changes promoted by inositol hexakisphosphate are largely dependent on spin state of the iron; tertiary and quaternary changes are not necessarily linked; structures induced by polyanions can be mixed forms that are neither T-state nor R-state.

Animals↗
PubMed1985

Conformational changes induced by polyanions in haemoglobin from Camelus dromedarius. Circular-dichroism study on the oxy derivative.

The c.d. spectrum of oxyhaemoglobin from Camelus dromedarius is significantly affected by the presence of inositol hexakisphosphate. Correlation with O2-binding measurements shows that these dichroic changes parallel the functional properties of the protein. The optical modifications suggest that, in contrast with human haemoglobin, the conformational changes induced by inositol hexakisphosphate on dromedary oxyhaemoglobin are mainly attributable to a local change of the tertiary structure reminiscent of that of the deoxy derivative, the quaternary conformation seeming to be almost unaffected. The results provide direct evidence of the existence on the protein of two distinct sites for polyanions.

Animals↗
PubMed1984

Oxygen binding to Octolasium complanatum erythrocruorin. Modulation of homo- and heterotropic interactions by cations.

The functional properties of erythrocruorin from Octolasium complanatum (a common earthworm of Central Italy) have been characterized in great detail. Special attention has been given to the reciprocal effects of the various ligands, namely oxygen, cations and protons. The data obtained under a variety of experimental conditions bring out the dominant role played by cations in the modulation of both homotropic and heterotropic interactions. In this respect, the most interesting observation concerns the unusual interplay between protons and cations that occurs in this erythrocruorin, the first respiratory pigment in which the Bohr effect is due totally to the O2-linked binding of an allosteric effector. The oxygen binding data collected under the various experimental conditions have been analyzed in terms of a modified two-state model, which takes into account the fact that allosteric effectors may also influence the ligand binding properties of the state that they stabilize. The analysis shows that the number of interacting sites necessary for the observed co-operativity in O2 binding is much smaller than the number of heme groups carried by the whole molecule, in accordance with previous findings on hemocyanins, the other class of giant respiratory pigments. Moreover, the analysis indicates that the dimensions of these "functional constellations" are under the control of allosteric effectors.

Allosteric Regulation↗
PubMed1984

Redox potentials of normal and SH(beta 93)-modified human hemoglobin. Effect of pH and D-glycerate 2,3-bisphosphate.

The effect of pH and D-glycerate 2,3-bisphosphate on the oxidation-reduction equilibria of human hemoglobin, normal and modified at the SH(beta 93) groups by reaction with cystine dimethylester has been studied with the aim of comparing the redox equilibria with the oxygen equilibria previously studied under the same experimental conditions. Analysis of data has shown that: (i) the same groups are involved in the oxygenation and oxidation alkaline Bohr effect; (ii) the pK values of these groups are lower in met-hemoglobin than in oxyhemoglobin; (iii) the difference in the affinity for D-glycerate 2,3-bisphosphate of ferric and deoxyhemoglobin is pH-independent in the normal protein, while significantly decreasing with a decrease in pH in the modified protein; (iv) the high cooperativity of the redox process (also occurring at acid pH) and the reduced oxidation Bohr effect observed in the modified protein are consistent with the destabilization of the T structure induced by the bulky reagent bound to the SH(beta 93) groups of the protein.

2,3-Diphosphoglycerate↗
PubMed1983

A circular dichroism study of the proton-linked transition in the carbomonoxy derivative of the hemoglobin component IV from trout.

Near ultraviolet and visible circular dichroism spectra of the carbomonoxy derivative of the hemoglobin component IV from trout Salmo irideus are pH-dependent in the range 6.2-7.8, and are affected by the presence of inositol hexaphosphate at pH 6.2. On the basis of previous studies, the spectral changes observed can be associated to the pH-dependent R4 leads to T4 transition occurring in the liganded protein. The CD spectra above 500 nm at low pH can be interpreted as due to release of the heme asymmetry in the T liganded conformation, in agreement with the movement of the iron toward the proximal histidine.

Allosteric Regulation↗
PubMed1982

Studies on Scapharca hemoglobins. Properties of the dimeric protein reconstituted with Fe- or Co-porphyrin.

A native globin from the dimeric hemoglobin, hemoglobin I, of the mollusc Scapharca inaequivalvis has been obtained with the acid-acetone method. The globin has a lower sedimentation coefficient than the native protein at neutral pH; its reconstitution product with natural heme has the same physicochemical and functional properties as the native protein. proto- and meso-cobalt hemoglobin I have been prepared and characterized. proto-Cobalt hemoglobin I binds oxygen reversibly with a lower affinity and a lower cooperativity than native hemoglobin I; thus, the changes in the functional properties brought about by substitution of iron with cobalt are similar to those observed in human hemoglobin A. The EPR spectra of deoxy-proto-cobalt hemoglobin I and of the photolysis product of oxy-meso-cobalt hemoglobin I indicate that two histidine residues are the apical heme ligands. The broad signal at g = 2.38 in deoxy-proto-cobalt hemoglobin I points to a constrained structure of the heme site in this derivative which results from a distorted coordination of the hindered proximal histidine. A similar structure has been proposed previously for the alpha chains in deoxy-cobalt hemoglobin A.

Apoproteins↗
PubMed1982

Circular-dichroism study of the interaction of aspartate-aminotransferase isoenzymes with a coenzyme analog.

The interaction between a coenzyme derivative, 4'-N-(2,4-dinitro-5-fluorophenyl)-pyridoxamine 5'-phosphate, and the apoenzyme of cytoplasmic and mitochondrial aspartate aminotransferase, was studied by circular dichroism. The specific complexes initially formed were characterized by their circular dichroic spectra. The spectra indicate that the complex is very probably the same for the two isoenzymes. In contrast the spectra recorded during further reaction, in agreement with previous results, monitor different reaction paths and characterize the irreversible labeling at the active site of the cytoplasmic enzyme and regeneration of pyridoxal 5'-phosphate in the mitochondrial enzyme. By following circular dichroic changes in the mitochondrial enzyme, initial kinetic characterization of the cleavage of 4'-N-(2,4-dinitro-5-fluorophenyl)-pyridoxamine 5'-phosphate to form pyridoxal 5'-phosphate at the active site, is provided.

Animals↗
PubMed1981

Evidence for multiple differing cation binding sites on hemoglobin A and S.

Some divalent cations (at 10(-3)M concentration) have been shown to act as oxygen-linked ligands. The effect of Ca++ (only on hemoglobin S) and Zn++ (on normal hemoglobin and hemoglobin S) can be traced to a preferential binding of these cations to the oxy- conformation of the proteins; the reverse is true for Ni++, since its presence reduces the oxygen affinity of both hemoglobins. The results suggest: i) the existence of at least two different cation binding sites on hemoglobin; ii) the replacement of glutamic acid by valine in hemoglobins S introduces sufficient structural modification to form a new cation binding site.

Binding Sites↗