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Biomedical subjects

K Dill

Publications and source records attributed to K Dill.

At least 37 records · Page 2Linked to original sources
PubMed1989

The interaction of phenyldichloroarsine with erythrocytes.

The purpose of the study was to identify binding sites of organic arsenic in the erythrocyte and to explain species differences in binding. Washed erythrocytes were exposed to graded concentrations of [U-14C]phenyldichloroarsine (PDA) in phosphate-buffered saline containing 0.1% glucose and 0.1% bovine serum albumin. At low PDA concentrations, all cells bound the arsenical rapidly (within 10 min) and quantitatively. Human, pig, hamster, guinea pig, and mouse erythrocytes approached saturation at 0.02-0.3 mumol PDA/10(9) cells, depending on the species. Saturation points correlated well with each respective species' erythrocyte glutathione content. In contrast, rat erythrocytes showed no sign of saturation at PDA loads as high as 3.0 mumol/10(9) cells. Hemolysates of PDA-treated erythrocytes were subjected to Sephadex G-75 gel filtration chromatography. 14C from rat hemolysate was distributed between the hemoglobin and small molecular weight (glutathione-containing) fractions. In all other species, the 14C eluted almost exclusively with the glutathione-containing fractions. In equilibrium dialysis experiments, human hemoglobin did not bind PDA, whereas rat hemoglobin bound 2 PDA/mol with Kd approximately 5 microM. In conclusion, glutathione is the principal binding site of phenyldichloroarsine in erythrocytes. In most species, the arsenical does not bind to hemoglobin, even though it has free (titratable) sulfhydryls considerably in excess of the glutathione concentration. In rat erythrocytes, phenlydichloroarsine binds both to glutathione and to hemoglobin. Arsenical binding by rat hemoglobin is presumably due to the unique location of the extra titratable cysteine in that protein.

Animals↗
PubMed1988

13C-nuclear magnetic resonance study of glycophorins AM and AN modified with various pyrylium salts.

The environment of the N-terminal amino groups of glycophorins AM and AN has been studied using 13C-NMR spectroscopy and pyrylium salts as amino-blocking agents. The extent of amino blocking was monitored by 13C-reductive methylation of the residual free amino groups. The pyrylium ions reacted with the N-terminal amino groups of the two glycophorins at almost identical rates, which is thought to indicate that the overriding steric bulk of the pyrylium salt may determine the rate of the reaction. The difference in the rates of modification of lysine residues of glycophorins AM) and AN by the pyrylium ions did indicate that there may exist an environmental difference around the lysine residues between the two glycophorins. This environmental difference may result from solution aggregation of the glycophorin A molecules or from some differences in the pKa values of the five lysine residues found in glycophorins AM and AN.

Chemical Phenomena↗
PubMed1987

One-dimensional and two-dimensional nuclear magnetic resonance studies of the reaction of phenyldichloroarsine with glutathione.

14C-labeled phenyldichloroarsine (PDA) enters the red blood cell and forms a 1:2 adduct with intracellular glutathione. Upon gel filtration of the hemolysate, [14C]PDA was recovered with the glutathione-containing fractions. One-dimensional and two-dimensional nuclear magnetic resonance spectroscopy were used to confirm the structure of the adduct and elucidate its stereochemistry, stability, and reactivity.

Arsenicals↗
PubMed1986

Possible role of the carbohydrate residues on the structure of the N-terminus of glycophorin AM.

Natural-abundance 13C nuclear magnetic resonance (13C-n.m.r.) was used to study the effect of monoglycosylation on the structure and dynamics of a pentapeptide related to the N-terminus of glycophorin AM. The results of this study indicate that a single point of glycosylation, on the pentapeptide, can significantly affect its structure. Moreover, glycosylation of this pentapeptide also affects its dynamic motion in solution. This study further defines the role that the carbohydrate residue plays in determining the structure about the N-terminus of glycophorin AM.

Carbohydrate Sequence↗
PubMed1986

13C n.m.r. study of the structure and the metal ion binding sites of neuropeptides composed of L-Asp and L-Glu.

13C NMR spectral data are presented for a variety of possible neuropeptides composed of L-Asp, Ac-L-Asp, and L-Glu which contain alpha and beta peptide linkages. The data for the various compounds are compared to the data presented for Ac-Asp-Glu, a known neuropeptide, in order to gain structural information about the related compounds. Indications are that for compounds 1 and 5, the cis peptide bond configuration exists due to the interaction of zwitterionic species. This interaction appears to be eliminated when the beta peptide bonds are formed, as in the case of compounds 3 and 7. Spin-lattice relaxation times were used to confirm the structures. Electron-nuclear relaxation rates are also used to elucidate the metal ion binding sites of these species.

Aspartic Acid↗
PubMed1985

Structural, dynamic, and metal-ion binding studies of the core glycopeptides beta-D-Gal-(1----3)-alpha-D-GalNAc----Ser, Thr by 13C-N.m.r. spectroscopy.

13C-N.m.r. spectral data as well as spin-lattice relaxation times (T1 values) are presented for the core glycopeptides beta-D-Gal-(1----3)-alpha-D-GalNAc----Ser, Thr. The binding of Gd3+ to these model compounds containing N-terminal blocking groups and esterified carboxyl groups indicates that the disaccharide contains a rather weak, but unique, binding-site in the vicinity of C-2 of alpha-D-GalNAc (possibly involving N-2', the acetamido carbonyl group, O-3' and/or possibly the glycosidic oxygen atom (O-3)).

Gadolinium↗
PubMed1985

A 13C-methylation study of glycophorin A intact erythrocytes by 13C-NMR spectroscopy.

N-terminal N alpha-[13C]monomethylamino derivatives for the N-terminal serine and leucine residues of glycophorins AM and AN, respectively, were obtained by reductively 13C-methylating homozygous human erythrocytes (MM, NN). The 13C-labeled glycophorins, AM and AN, were then isolated. A unique structural state was observed in solution reductively 13C-methylated glycophorin AM that was not observed in glycophorin AM derived from 13C-methylated erythrocytes. We attribute this state to the fact that some of the glycophorin AM forms a head-to-head dimer when subjected to reductive 13 C-methylation in aqueous solution. The 13C chemical shift data and pH titration data for the N-terminal [13C]dimethylamino and [13C]monomethylamino groups of glycophorin AM and AN derived from reductively 13C-methylated erythrocytes were in agreement with the chemical shift and titration data previously obtained for the N-terminal [13C]dimethylamino groups of solution reductively 13C-methylated glycophorins and related glycopeptides and peptides and N-terminal [13C]monomethylamino groups of related glycopeptides and peptides.

Chemical Phenomena↗
PubMed1984

13C-NMR spectral study of reductively [13C]methylated glycophorin B.

Glycophorin BN was reductively [13C]methylated and the 13C chemical shift of the N-terminal [13C]dimethyl-leucine residue was monitored as a function of pH. These results were compared to the pH-dependent chemical shift studies of the N-terminal [13C]dimethylleucine residues of intact glycophorin AN and N-terminal glyco-octapeptide AN. The results indicate that the titration data for [13C]dimethylleucine of glycophorin BN more closely resembles the titration data observed for the [13C]dimethylleucine residue of the N-terminal glyco-octapeptide AN rather than for the [13C]dimethylleucine residue of intact glycophorin AN. Integration of the 13C resonances indicated that glycophorin BN contains 3-4 lysine residues.

Glycophorins↗
PubMed1983

13C-N.m.r.-spectral study of the mode of binding of Gd3+ to various glycopeptides.

Natural-abundance, 13C-n.m.r. spectroscopy was used to study the mode of binding of Gd3+ to mono-O-glycosylated L-serine and tripeptides variously composed of Gly and L-Thr. When the amino and carboxyl groups of the amino acid are not blocked, strong interaction of Gd3+ with them is observed; this is also readily apparent with some related, nonglycosylated peptides. When the amino and carboxyl groups of the amino acid are blocked, noticeable interaction of Gd3+ with the glycosidic oxygen atom (O-3) and O-2' for the glycopeptide containing alpha-D-Galp, and with O-3 and N-2' for the glycopeptide containing alpha-D-GalpNAc, is observed. Weak interactions are also possible with O-4' and O-6' of the glycosyl groups. Although the amino acids were protected, these metal ion-carbohydrate interactions may still be mediated, to some extent, by the acetyl protecting the amino group and by the ester group on the amino acid.

Gadolinium↗
PubMed1983

Possible role of the carbohydrate residues in the display of the MN blood group determinants by glycophorin A.

Heterozygous glycophorin AM,N and homozygous glycophorin AM were reductively methylated with 13C-enriched formaldehyde in the presence of cyanoborohydride. Total reductive methylation modified the five lysine residues, and the N-terminal amino acid residues (serine and leucine) of glycophorins AM and AN, respectively. The 13C resonances of the incorporated labels were monitored as a function of the degree of glycosylation of the glycoprotein. While minimal, if any, structural changes were observed near the N-terminal amino acid upon removal of alpha-D-N-acetylneuraminic acid residues, gross structural changes were observed when most of the oligosaccharide chains were removed. We also found that progressive methylation of the lysine residues of glycophorin AM may influence either the chemical shift of one of the nonequivalent methyl groups of the N alpha, N-[13C]dimethyl serine residue, or one of the two states of glycophorin AM.

Animals↗
PubMed1983

Specific 13C reductive methylation of glycophorin A. Possible relation of the N-terminal amino acid and the lysine residues to MN blood group specificities.

Heterozygous and homozygous glycophorin A were partially and fully reductively methylated with 13C-enriched formaldehyde in the presence of sodium cyanoborohydride. Total reductive methylation modified the five lysine residues (to produce N epsilon,N-[13C]dimethyl lysine) and the N-terminal amino acid residues (N alpha,N-[13C]dimethyl serine and leucine) of glycophorins AM and AN, respectively. 13C-NMR spectra of these species indicated that the 13C-enriched methyl carbons of the five lysyl derivatives all occur at 44.1 ppm downfield from Me4Si. Titration results indicate that the pK alpha of these methylated lysines is greater than 10. The chemical shift equivalent methyl resonances of the 13C-enriched methylated N-terminal Leu derivative were found to occur at 42.8 ppm downfield from Me4Si and exhibited a normal pH titration behavior (pK alpha approximately 7.4). The methyl resonances of the N alpha,N-[13C]dimethyl Ser derivative, on the other hand, were found to exhibit chemical shift nonequivalence, indicating rotational constraints about the C alpha-N bond. The linewidths of the two methyl resonances were also found to be considerably different; this phenomenon could be eliminated by running spectra of the sample (pH approximately 5.0) at elevated temperatures (75 degrees C). This result suggested that for the N alpha,N-[13C]dimethyl Ser derivative of glycophorin AM, hindered rotation must occur about one of the N alpha-13CH3 bonds. This structural difference at the N-terminal residue of glycophorins AM and AN may be related to the MN blood group determinants displayed by these related glycoproteins.

Amino Acid Sequence↗
PubMed1983

13C-N.M.R.-spectral study of the binding of Gd3+ to glycophorin.

Natural-abundance, 13C-n.m.r. spectroscopy was used to study the binding of Gd3+ to glycophorin, and also to the tetrasaccharides isolated from glycophorin after treatment of the glycoprotein with NaOH-NaBH4. Gd3+ binds to the tetrasaccharide (both in the isolated, reduced form and when still attached to the native glycoprotein), and, especially, to the alpha-NeuAc residues. In order to cause severe line-broadening of the 13C resonances of alpha-NeuAc, the ratios of the alpha-NeuAc residues of glycophorin, and of the isolated, reduced tetrasaccharide, to Gd3+ were much higher than that needed for causing similar broadening for 2-O-methyl-alpha-NeuAc-Gd3+ solutions. These results indicate that the other carbohydrate residues of the tetrasaccharide may be involved in the binding of Gd3+, producing a stronger metal-ion-binding effect.

Binding Sites↗
PubMed1982

Magnetic resonance study of glycophorin A-containing 13C-enriched methionines.

Methionine-81 and/or -8 of the transmembrane sialoglycoprotein, glycophorin A, have been specifically alkylated with 13CH3I to produce the sulfonium ion derivatives [S-[13C]methylmethionine-8]glycophorin A and [S-[13C]methylmethionine-8 and -81]glycophorin A. 13C NMR spectra of these species show that the resonances of the methyl groups of the modified glycophorins occur at 26.1 ppm downfield from Me4Si. A spin-lattice relaxation time of 0.4 s was observed for the 13C-enriched methyl resonances of the sulfonium ion derivatives of Met-8 and -81, which corresponds to an effective correlation time of less than 2 X 10 - 10 s. Demethylation of the 2 glycophorin A sulfonium ion species with 2-mercaptoethanol produces native glycophorin A which now has the epsilon-carbon of the methionine residue(s) 45% isotopically enriched. The epsilon-carbon of Met-8 was found to occur at 15.7 ppm downfield from Me4Si whereas the epsilon-carbon of Met-81 exhibited an unusual chemical shift of 2.0 ppm downfield from Me4Si. The spin-lattice relaxation time of both resonances was found to be approximately 0.3 s.

Carbon Isotopes↗