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C Turano

Publications and source records attributed to C Turano.

At least 37 records · Page 2Linked to original sources
PubMed1991

A reverse-phase HPLC method for cAMP phosphodiesterase activity.

A simple and fast method based on reverse-phase HPLC has been developed for measuring the activity of cAMP phosphodiesterase. It allows quantitation of product and substrate in less than 10 min. The sensitivity (1*10(-11) mol AMP), the accurate evaluation of nucleotides, the unequivocal analysis of product, and the reproducibility of the system, make this method suitable for the evaluation of cAMP phosphodiesterase in biological material, at different levels of purification, and also in kinetic studies.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1989

Glycosylated forms of nuclear lamins.

Chromatin and pore complex-lamina preparations were obtained from pig and chicken tissues, and their proteins were analysed by mono- and bidimensional electrophoresis. A glycosylated form of lamin A, recognized by concanavalin A, was shown to be present in at least 3 of the tissues examined. Glycosylation is suggested to be a further postsynthetic modification, besides phosphorylation and methylation, which can modify the properties of lamins.

Animals↗
PubMed1989

Tissue specificity of chromatin glycoproteins recognized by concanavalin A.

Chromatin glycoproteins recognized by Concanavalin A have been isolated from pig liver, kidney and heart by the use of immobilized lectin. Two groups of proteins differing in affinity for DNA have been analysed. Glycoproteins are mainly present in the group of proteins which are tightly bound to DNA. Mono and bidimensional electrophoretic patterns of total tightly bound proteins reveal a similarity among the three organs examined, while the corresponding patterns of the glycoproteins are typical for each organ. The tissue specificity of chromatin glycoproteins, together with their capability to interact not only with DNA but possibly also with other nuclear components, suggest a role for these proteins in the mechanism of genome expression.

Animals↗
PubMed1988

Calorimetric analysis of sodium dodecylsulfate-chromatin interaction.

Microcalorimetric titrations of whole chromatin and histones with sodium dodecylsulfate were performed at pH 7 and 25 degrees C. Enthalpy variations at low detergent concentration (less than 0.02%) are much more negative for histones than for chromatin. At 0.065% sodium dodecylsulfate the difference between the two curves becomes constant and, after correction for monomerization effects, amounts to +130 kcal/mol of nucleosomal unit. Core particles show heat effects similar to those of histones. These findings suggest that the chromatin structure is not stabilized exclusively by electrostatic interactions and that hydrogen bonds responsible for the additional stability may be contributed by non histone chromatin proteins.

Animals↗
PubMed1986

The influence of ionic strength on the binding of a water soluble porphyrin to nucleic acids.

The ionic strength dependences of the binding of tetrakis (4-N-methylpyridyl)porphine (H2TMpyP) to poly(dG-dC) and calf thymus DNA have been determined. For the former system the results are typical of other intercalators, i.e., a plot of log K vs log [Na+] is linear albeit with a slope which suggests that the "effective charge" of the porphyrin is closer to two than the formal charge of +4. For calf thymus DNA, the binding profile is not completely compatible with the predictions of condensation theory. Whereas the avidity of binding does decrease with increasing [Na+] as predicted, of greater interest is the relocation of the porphyrin from GC-rich regions to AT-rich regions as the ionic strength increases.

Animals↗
PubMed1986

Thiol proteins in chromatin.

Total half-cystine residues in proteins of pig liver chromatin have been measured. About half of them are present in the reduced state. Thiol groups of non-histone chromatin proteins, which amount to about 40 nmol/mg of protein, are preferentially located in chromatin fragments which are more easily solubilised either by DNAse I or by DNAse II. The data obtained are compatible with an involvement of SH and SS groups in chromatin structure and function.

Amino Acids↗
PubMed1985

Specific labeling of cytosolic and mitochondrial aspartate aminotransferases.

The apoisozymes of cytosolic and mitochondrial aspartate aminotransferase are both irreversibly inhibited by alpha-N-fluorodinitrophenyl-beta-N-phosphopyridoxyldiaminopropi onate, an affinity-labeling reagent analog of the coenzyme. Analysis of the modified peptides shows that the active-site Lys-258, which in the holoenzyme binds the coenzyme pyridoxal 5'-phosphate, is labeled in both isozymes. Comparison with the results obtained using the parent compound 4'-N-fluorodinitrophenylpyridoxamine 5'-phosphate, which labels only the cytosolic enzyme, provides information about differences in active-site reactivity and geometry. Labeling external to the active site occurs in both isozymes. In the cytosolic enzyme the very reactive Cys-45 is modified, in the mitochondrial enzyme the surface residue Lys-342 reveals a peculiar reactivity.

Affinity Labels↗
PubMed1984

The interaction of bisbenzimide with DNA.

The interaction of bisbenzimide (Hoechst 33258) with DNA has been studied by microcalorimetry, fluorimetry and fluorescence polarization measurements. The binding does not involve intercalation, takes place in at least two modes and occurs in the major groove at low dye/DNA molar ratio (first mode of binding), in agreement with previous proposals by other Authors. Furthermore it has been shown that i) at high dye/DNA molar ratio the binding (second mode) occurs in both grooves; ii) at pH 7.0 the protonation of a dye nitrogen accompanies the binding in both modes; iii) the quantum yield of the dye-DNA complex is sensitive to the DNA conformation change caused by an increase in ionic strength. Basically the same type of binding takes place on chromatin, as detected by calorimetry. However, at high dye/DNA molar ratio about 30% of DNA is not accessible to the dye. The decreased accessibility appears to be mainly related to the nucleosomal structure of chromatin.

Animals↗
PubMed1982

A calorimetric study of the interaction of pyridoxal 5'-phosphate with aspartate apoaminotransferase and model compounds.

Schiff base formation between pyridoxal 5'-phosphate and model compounds of increasing complexity (i.e. L-valine and poly-L-lysine) and between pyridoxal 5'-phosphate and the apoenzyme of aspartate aminotransferase has been analyzed by microcalorimetric methods. The apparent pKa values and protonation enthalpy values for the relevant groups ionizing in the pH 4-9 range have been determined for the Schiff bases of L-valine and of poly-L-lysine. Upon Schiff base formation, the only noticeable change is the lowering of the ring nitrogen pK, accompanied by an increase of the relative delta H. In the poly-L-lysine Schiff base, however, the delta H relative to the protonation of the phosphate dianion becomes more negative. This behavior suggests a multiple interaction between the polymer and the ligand. The intrinsic heat of formation is small (congruent to -1 kcal/mol), of the same order of magnitude for both Schiff bases, and appears to be independent of the nature of the aminic reagent. The heat of reaction of pyridoxal 5'-phosphate with aspartate apoaminotransferase has been determined in the pH 6.2-8.8 range at 19 degrees C and at 25 degrees C. Each isotherm is characterized by a lack of proton evolution, a result that is unexpected on the basis of the known pK values of the reagents, and by a sharp pH dependence of the enthalpy change. Moreover, comparison of the two isotherms allows: (a) detection of a protonation-dependent effect (pK 7.5 at 25 degrees C), (b) exclusion of a preferential binding of the coenzyme to the apoenzyme in a particular ionization state; and (c) suggestion of a tightening of the protein molecule upon holoenzyme formation.

Animals↗
PubMed1981

Tightly bound non-histone proteins in nucleosomes from pig-liver chromatin.

Core particles prepared by micrococcal nuclease digestion of pig liver chromatin have been adsorbed on hydroxyapatite and dissociated by gradual increase in ionic strength and finally by urea and guanidine. By this method non-histone proteins have been found to be associated with the core particles. Proteins tightly bound to the core particle DNA (i.e. dissociated only by urea and guanidine) have also been found: these are proteins with a limited heterogeneity, with respect to their molecular weights, since only six components are present with molecular weights ranging from 71000 to 20000. They show, furthermore, a peculiar amino acid composition. Other tightly bound proteins have been shown to be present only in the spacer regions. The existence of two different classes of tightly bound proteins probably reflects different modes of binding to the DNA, which are compatible or incompatible, respectively, with the simultaneous binding of the histone octamer.

Animals↗
PubMed1980

Different reactivity of mitochondrial and cytoplasmic aspartate aminotransferases toward an affinity labeling reagent analog of the coenzyme.

The two isoenzymes of aspartate aminotransferase from pig heart have been reacted with a derivative of the coenzyme, 4'-N-(2,4-dinitro-5-fluorophenyl) pyridoxamine-5'-phosphate, which is a potential affinity labeling reagent. The derivative has a great affinity for both isoapoenzymes. In the cytosolic isoenzyme, the reversible binding is followed by a covalent labeling of the epsilon amino group of lysine 258, which usually forms an aldimine bond with pyridoxal-5'-phosphate. In the mitochondrial isoenzyme, no labeling occurs at the active site. The different reactivity indicates that a small but definite difference exists in the geometry of the two active sites. In the cytosolic isoenzyme also a sulfhydryl group outside the active site region, namely cysteine 45, reacts, but not by an affinity labeling mechanism. In both isoenzymes, the reversibly bound reagent slowly undergoes a splitting reaction by which pyridoxal-5'-phosphate is regenerated and activity re-established; the rate of this reaction is not fast enough to impaire the labeling potential of the reagent.

Affinity Labels↗