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Biomedical subjects

G Colonna

Publications and source records attributed to G Colonna.

At least 73 records · Page 4Linked to original sources
PubMed1983

Structural and functional aspects of the heart ventricle myoglobin of bluefin tuna.

The heart ventricle myoglobin of bluefin tuna has been purified to an apparent homogeneity. The amino acid analysis has revealed only a limited number of substitutions between the myoglobins of yellowfin and bluefin tuna. The alpha-helix content of tuna myoglobin has been found considerably lower than that of mammalian myoglobin. No correlation has been discovered between the conformational stability and alpha-helix content. Denaturation experiments have shown that the whole structure of tuna myoglobin results from the interaction of two structural units which represent the product of independent folding processes. The structure of tuna myoglobin has been found more open and disorganized than that of sperm whale. This result has been related to the low content of electrostatic interactions and explained in terms of evolutive adaptations.

Amino Acid Sequence↗
PubMed1983

Non-invasive quantification of aortic regurgitation by Doppler echocardiography.

This study was undertaken to assess the contribution of Doppler echocardiography to the quantification of aortic valve regurgitation. Ultrasound examination was performed by recording aortic arch blood flow from the suprasternal notch. A non-invasive index of valve regurgitation was obtained by calculating the ratio between the maximal amplitude of forward flow during systole and the amplitude of retrograde flow during diastole measured at the onset of the R wave of the electrocardiogram. This index was compared with semiquantitative data derived from supravalvular aortography in 93 patients. In pure aortic regurgitation (67 patients) the results showed a high correlation coefficient between Doppler and angiographic estimates. In cases of associated aortic valve stenosis there were problems in the accurate estimation of systolic blood flow which led to global overestimation in general of the degree of regurgitation and considerable lack of precision in individual patients. But in general Doppler echocardiography appeared to be a successful technique to quantify pure aortic regurgitation.

Adolescent↗
PubMed1983

Quantitative assessment of tricuspid regurgitation using pulsed Doppler echocardiography.

Tricuspid valve regurgitation was assessed quantitatively by measuring blood flow velocity in the vena cava using a pulsed Doppler velocimeter. A non-invasive index of regurgitation was obtained by calculating the ratio between the maximum amplitudes of the systolic and diastolic components of the velocity curves. The index was compared with the angiographic grading of regurgitation in 70 patients after right heart catheterisation; the results were closely correlated. Using the Doppler index the differences between the groups defined according to their angiographic grade were significant. Thus measurement of blood flow velocity in the vena cava appears to quantify accurately the severity of tricuspid regurgitation.

Adolescent↗
PubMed1982

Purification and molecular properties of rabbit lung indolamine N-methyltransferase.

Indolamine N-methyltransferase (INMT) has been purified to an apparent homogeneity from rabbit lung, and some of its catalytic and physicochemical properties have been examined. The enzyme is a monomeric protein with a molecular weight of 31,500 +/- 1000, a molecular Stokes radius of 21.5 A, and a diffusion coefficient of 8.7 X 10(-7) cm2/s. The frictional ratio of the native enzyme (1.05) suggests that the shape of the molecule is nearly spherical. Denaturation experiments performed with increasing concentrations of guanidine hydrochloride (Gdn-HCl) at neural pH indicated that the active site of the enzyme was destroyed by a structural rearrangement of the protein molecule without large change in its size and shape. The final state reached in 6.0 M Gdn . HCl seemed to correspond to a disulfide cross-linked randomly coiled polypeptide. Full normalization of the fluorescent parameter was attained only in the presence of 0.1 M beta-mercaptoethanol. A structural rearrangement has been observed upon acidification of INMT from pH 7.0 to pH 2.0. At pH 4.5, most of the peptide backbone appeared to be unorganized, but further acidification to pH 2.0 produced a reorganization of protein structure which became able to bind 8-anilino-1-naphthalenesulfonate. The data support the hypothesis that the enzyme structure results from the close package of organized regions joined by structureless segments.

Animals↗
PubMed1982

Heme and cysteine microenvironments of tuna apomyoglobin. Evidence of two independent unfolding regions.

The heme and cysteine microenvironments of bluefin tuna apomyoglobin have been investigated by examining the fluorescence properties of two extrinsic chromophores, i.e., ANS and 1,5-AEDANS. 1,5-AEDANS was covalently bound to the single cysteine residue found in the primary structure of tuna apomyoglobin. Recombination experiments with hemin showed that tuna apomyoglobin does not bind 1,5-AEDANS in the same binding site of the heme, although the fluorescence properties of the covalently bound 1,5-AEDANS strongly suggest that the dye is embedded in a rather nonpolar microenvironment. ANS was selected because of its ability to bind the apomyoglobin in the same nonpolar moiety of the heme. Acidification of apoMb--AEDANS to pH 3.0 produced an increase of 1,5-AEDANS fluorescence intensity and a shift of its emission maximum from 475 to 470 nm. In the same pH range apomyoglobin lost its ability to bind ANS. Two independent transitions were observed with increasing concentrations of guanidine. Low guanidine concentration (less than 1.0 M) unfolded the heme binding site as indicated by the disappearance of ANS fluorescence, whereas higher denaturant concentration was required to produce full normalization of 1,5-AEDANS emission spectrum.

Animals↗
PubMed1982

[Non-invasive study of aortic insufficiency by Doppler echocardiography].

The possibilities of diagnosis and quantification of aortic regurgitation by pulsed Doppler analysis of blood flow in the aortic arch were examined in 60 patients aged between 9 and 67 years old. Aortic flow curves were recorded from the suprasternal area with the sample volume positioned at the junction of the horizontal part of the aortic arch and the descending aorta. Normal flow curves are characterised by an anterograde systolic wave with a brief proto-diastolic reflux. In aortic regurgitation holodiastolic reflux is observed. An index of regurgitation may be calculated from the ratio of the amplitude of end diastolic reflux measured on the R wave of th ECG and the maximal amplitude of anterograde systolic flow. This ratio eliminates the factor related to the incident angle between the ultrasound beam and the direction of blood flow. The values of this ratio were compared to the semi quantitative assessment of aortic regurgitation from ascending aortic angiography. The only false negatives were observed in patients with negligible regurgitation (grade I). One false positive result was obtained in a patient in whom it was difficult to obtain the recording and in whom the value of the ratio was very low (0,02). Global specificity was 91 p. 100 and sensitivity was 82 p. 100. The sensitivity for average or severe regurgitation was 100 p. 100. The correlation coefficient between the Doppler index or regurgitation and the semi quantitative angiographic estimation was 0,69. In patients with pure aortic regurgitation the correlation reached 0,85 (p less than 0,001). The differences between the different groups then became highly significant.

Adolescent↗
PubMed1981

Tryptophanyl fluorescence heterogeneity of apomyoglobins. Correlation with the presence of two distinct structural domains.

The individual fluorescence of the two tryptophan residues (Trp-7 and Trp-14) of mammalian apomyoglobins has been resolved by comparing the fluorescence properties of these proteins to those of bluefin tuna apomyoglobin, which contains only Trp-14. The two tryptophan residues have been found to have different emission maxima, i.e., 321 for Trp-14 and 333 for Trp-7. The fluorescence of Trp-14 depends on the protonation of a sterically related histidyl residue in the pH range between 8.3 and 5.6, where no conformational change was detected. This residue has been identified as His-119. At pH 8.3 the quantum yield of Trp-7 is lower than that of Trp-14. An increase of the fluorescence intensity of Trp-7 occurs when the heme binding site of apomyoglobin is destroyed by acid or a low concentration of guanidine hydrochloride. An independent unfolding of the N-terminal district of the apomyoglobin molecular occurs on increasing the guanidine concentration. The two distinct structural transitions have been discussed in terms of two domains of tertiary structure.

Animals↗
PubMed1979

The structure and stability of human plasma cold-insoluble globulin.

The molecular properties of cold-insoluble globulin have been investigated by velocity centrifugation, circular dichroism, and fluorescence at neutral and alkaline pH. The stability of the protein to thermal and guanidine hydrochloride has been evaluated under both conditions. The close parallelism between the properties of cold-insoluble globulin and those of the cell surface protein (fibronectin) serve to establish the essential identity of the structures of the two proteins derived from different sources. It is suggested that the cold-insoluble globulin is composed of several domains connected by flexible polypeptide segments. The large increase in the frictional ratio observed between pH 7.0 and 11.0 can be explained by an expansion of the flexible segments without significant change in the domains. These domains are stable to about 55 degrees C at pH 7.0 but only to about 40 degrees C at pH 11.0.

Circular Dichroism↗
PubMed1978

The stability of cell surface protein to surfactants and denaturants.

The effects of several denaturants and detergents on the structure and stability of cell surface protein have been evaluated by circular dichroism and fluorescence measurements. Cell surface protein undergoes a single broad transition in both urea and guanidinium chloride. Although guanidinium chloride is twice as effective as urea on a molar basis, both appear to eliminate all of the organized structure present in the native molecule. Nonionic surfactants and lysolecithin have little effect on cell surface protein. However, sodium dodecyl sulfate increases the alpha helical content and cetyltrimethylammonium bromide increases the beta structure of cell surface protein. The reorganization of the polypeptide backbone requires the loss of certain restraints imposed by tertiary interactions as evidenced by a decrease in ellipticity in the far ultraviolet and in the polarization of tryptophanyl fluorescence. These results along with the data of a previous paper (Alexander, S. S., Jr., Colonna, G., Yamada, K. M., Pastan, I., and Edelhoch, H. (1978) J. Biol. Chem. 253, 5820--5824) suggest the presence of structural domains distributed along the flexible polypeptide chain of cell surface protein.

Animals↗
PubMed1978

Second-derivative spectroscopy of proteins. A method for the quantitative determination of aromatic amino acids in proteins.

Second derivative spectroscopy has been used to resolve the complex protein spectrum in the near-ultraviolet region and the contributions of the three aromatic chromophores have been evaluated. A method for the direct quantitative determination of phenylalanine and tryptophan in proteins has been carried out. Phenylalanine determination has been carried out in the spectral region between 250 and 265 nm, where there are no significant contributions from other aromatic chromophores. Tryptophan determination has been performed in the 290-295-nm region and the experimental values have been corrected for the presence of tyrosine. The results obtained on 10 highly purified proteins have been found in good agreement with those obtained from sequence analysis.

Phenylalanine↗
PubMed1978

Molecular properties of a major cell surface protein from chick embryo fibroblasts.

The molecular structure of chick embryo fibroblast cell surface protein has been investigated by ultracentrifugation, circular dichroism, and fluorescence. Most measurements were restricted to alkaline solutions because of the limited solubility of this protein at more neutral pH values. A very high frictional ratio for the protein suggests an asymmetric structure. However, there are elements of organized structure since typical thermal transition curves were found by several methods. Consequently, a model in which ordered domains are connected by flexible polypeptide chains seems to account for all the hydrodynamic and optical data.

Animals↗
PubMed1978

The effect of evolution on homologous proteins: a comparison between the chromophore microenvironments of Italian water buffalo (Bos bubalus, L.) and sperm whale apomyoglobin.

The perturbing effect of guanidium hydrochloride and pH on the molecular structure of water buffalo apomyoglobin has been investigated by circular dichroism in the far and near ultraviolet and by fluorescence. In the wavelength region between 320 and 260 nm the circular dichroic spectrum of the globin is highly structured and the contributions of the aromatic chromophores have been resolved. Buffalo apomyoglobin undergoes a structural transition at neutral pH which involves elements of the secondary and tertiary structure, as indicated by changes of dichroic activity of the peptide and aromatic chromophores and the fluorescence of the two tryptophanyl residues. The possibility of charge-transfer complex between indole and imidazole is discussed. A major structural transition with abrupt unfolding takes place in the pH region between 5.6 and 4.3. Below pH 4.3 the peptide helical residues, which survive the acid transition, appear to be resistent to further acidification to pH 2.0 while tryptophanyl emission is quenched and shifted to longer wavelengths. A structural transition occurs also in alkali above pH 10, which has been detected by the same techniques. The relationships between buffalo and sperm whale apomyoglobin are discussed.

Animals↗
PubMed1978

Amino acid composition and physico-chemical properties of bluefin tuna (Thunnus thynnus) myoglobin.

1. The heart ventricle myoglobin of Atlantic bluefin tuna has been purified and its amino acid composition has been determined. 2. The perturbing effect of guanidine hydrochloride on the molecular structure of tuna ferrimyoglobin and its corresponding apoprotein has been investigated by Soret absorbance and ultraviolet fluorescence. 3. The conformation-free energy of unfolding delta G0 has been calculated by thermodynamic treatments of the data concerning guanidine unfolding. 4. The results have been compared with other known myoglobins, particularly those of yellowfin tuna.

Amino Acids↗