Biomedical subjects

T Drakenberg

Publications and source records attributed to T Drakenberg.

124 records · Page 7Linked to original sources

PubMed1984

Reaction of histamine with o-phthalaldehyde: isolation and analysis of the fluorophore.

Histamine reacts with o-phthalaldehyde (OPT) to form colored as well as highly fluorescent derivatives. After large-scale synthesis, the chromophores were eliminated and the pale yellowish fluorophore was isolated by a series of chromatographic steps, followed by semipreparative high-performance liquid chromatography. Structural analysis of the fluorophore was performed using NMR and mass spectrometry. The isolated fluorophore was found to be a condensation product of 2 molecules of OPT and 1 molecule of histamine, arranged to form a complex dihydrophenantroline derivative with a quaternary amine group. The molecular weight of its chloride salt was 379.5. It was shown that the fluorophore exists in aqueous milieu as an equilibrium between three isomers.

Aldehydes↗

PubMed1983

Calcium and cadmium binding to troponin C. Evidence for cooperativity.

Proton NMR is used to compare the structural changes induced in bovine cardiac troponin C on binding of cadmium and calcium ions. The same spectral changes are observed for both ion species. The rate of the conformational changes associated with cadmium binding to the two high-affinity sites is slow, that associated with cadmium ions binding to the low-affinity site is high. 113Cd-NMR spectra of cardiac troponin C feature two signals interpreted as due to cadmium ions bound to the strong sites. Strong arguments are given in favour of cooperativity in binding of the first two cadmium or calcium ions to cardiac and skeletal muscle troponin C.

Allosteric Regulation↗

PubMed1983

A 113Cd and 1H NMR study of the interaction of calmodulin with D600, trifluoperazine and some other hydrophobic drugs.

The interaction of calmodulin with D600 (a methoxy derivative of verapamil), trifluoperazine and some other drugs was studied by 113Cd and 1H NMR. All four cation binding sites of calmodulin were found to be affected by the binding of the drugs to calmodulin. The physiologically active and inactive forms of felodipine were found to give qualitatively the same changes in the 113Cd NMR spectra of calmodulin. The interpretation of this observation in terms of the physiological relevance of the binding to calmodulin is discussed. The binding constants for the two strongly bound trifluoperazine molecules were found to differ by one or two orders of magnitude. A competition study showed that trifluoperazine replaces D600 from at least one binding site on calmodulin. Moreover the binding of D600 was found to be calcium dependent. The data indicate that two calcium ions bound to calmodulin are sufficient to render the binding site(s) accessible for D600.

Animals↗

PubMed1983

beta-Hydroxyaspartic acid in vitamin K-dependent protein C.

Previous work has shown that the light chain of protein C, an anticoagulant plasma protein, contains an unusual amino acid [Fernlund, P. & Stenflo, J. (1982) J. Biol. Chem. 257, 12170-12179]. To determine the structure of this amino acid a heptapeptide, CMCys-Ile-X-Gly-Leu-Gly-Gly (residues 69-75 in the light chain), was isolated from enzymatic digests of the light chain. According to automatic Edman sequence analysis, 1H NMR spectroscopy, and mass spectrometry the heptapeptide had beta-hydroxyaspartic acid in its third position, which corresponds to position 71 in the light chain of protein C. Analysis of acid and aminopeptidase M hydrolysates of the heptapeptide showed the beta-hydroxyaspartic acid to be the erythro form. Acid hydrolysis of protein C released approximately equal to 1 mol of beta-hydroxyaspartic acid per mol of protein. The function of this amino acid, which, to the best of our knowledge, has not been found previously in proteins, is unknown.

Amino Acid Sequence↗

PubMed1982

Characterization of the Ca2+ binding sites of calmodulin from bovine testis using 43Ca and 113Cd NMR.

The exchange rates of Ca2+ ions to the two classes of sites on calmodulin have been determined from the temperature dependence of the 43Ca NMR line width. The exchange rates were found to differ by a factor of about 40 at room temperature. The apparent pK value for one of these classes was estimated from the pH dependence of the 43Ca line width. The pK values of the two-high-affinity sites were found to differ about 0.1-1. Zn2+ ions were shown to bind to calmodulin and affect the exchange rates of Ca2+ and Cd2+ for all four sites.

Animals↗

PubMed1982

Observation of the 43Ca NMR signals from Ca2+ ions bound to calmodulin, parvalbumin, and troponin C.

Through the combined use of isotopically enriched 43Ca2+, high magnetic field, Fourier transform techniques, and a high-performance probe, the 43Ca NMR signals from ions bound to three proteins have been observed. These proteins are calmodulin, parvalbumin, and troponin C. Both longitudinal (1/T1) and transverse (1/T2) relaxation rates were measured for protein-bound 43Ca2+, and the quadrupole coupling constant and correlation time were obtained from these data. The calculated correlation times are in good agreement with the rotational correlation time for the entire protein molecules, which indicates that the Ca2+-binding sites are relatively rigid.

Animals↗

PubMed1981