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C Kreutzfeldt

Publications and source records attributed to C Kreutzfeldt.

10 recordsLinked to original sources
PubMed1988

The anion-transport inhibitor H2DIDS cross-links hemoglobin interdimerically and enhances oxygen unloading.

Human hemoglobin treated with equal concentrations of the anion-transport inhibitor H2DIDS produces a right shift in the oxygen dissociation curve. concomitantly, the Hill coefficient is reduced from n = 2.7 to 2.1. When higher concentrations of H2DIDS are applied (H2DIDS: hemoglobin = 5:0.5 mM), the Hill coefficient decreases further to 1.5 and the oxygen dissociation curve of hemoglobin is shifted slightly to the left of the control. Similar results were also obtained with DIDS instead of H2DIDS. SDS-PAGE shows that H2DIDS cross-links hemoglobin monomers mainly into dimers. Cross-linking is more effective under anaerobic conditions. With tritiated H2DIDS the larger part of the radioactivity is found in the dimer position of hemoglobin. Separation of the alpha and beta units of hemoglobin reacted with tritiated H2DIDS demonstrated a stoichiometry of 2.2 and 2.4 molecules H2DIDS per molecule alpha and beta unit hemoglobin, leading to about 8-9 H2DIDS molecules per native hemoglobin. The right shift produced in the hemoglobin oxygen dissociation curve and the cross-linking of monomers into dimers, especially under anaerobic condition, suggest that H2DIDS can also react with those amino groups of hemoglobin which are involved in 2,3-DPG binding. A comparison of H2DIDS, DIDS and 2,3-DPG at three different concentrations close to the hemoglobin concentration revealed a concentration dependent right shift in the oxygen dissociation curve with the order of potency 2,3-DPG greater than H2DIDS greater than DIDS. The Hill coefficients (n) at the three concentrations of 2,3-DPG demonstrated no changes, but H2DIDS and DIDS reduced in a concentration-dependent manner the cooperativity of hemoglobin. Again, H2DIDS is more potent than DIDS, especially at the low concentration. These anion-transport inhibitors provide novel approaches to the exploration of hemoglobin function.

2,3-Diphosphoglycerate↗
PubMed1988

Immunological homologies between yeast ribosomal protein L2 and rat liver ribosomal proteins L4 and L24.

Polyclonal antibodies raised against ribosomal protein (r-protein) L2 of Schizosaccharomyces pombe were used to check for cross-reactions with total r-proteins of rat liver. Using this procedure, the rat liver r-proteins, L4 and L24, were identified as being immunologically related to yeast L2. In additional, homologies between rat liver L4 and L24 were detected. The possible implications for the regulation of r-protein synthesis are discussed.

Animals↗
PubMed1986

Immunological homologies between ribosomal proteins amongst lower eukaryotes.

Polyclonal antibodies were raised against the purified ribosomal proteins L1 and L2, the 5S rRNA binding protein L3, all from Saccharomyces cerevisiae, and against L1 and L2 from Schizosaccharomyces pombe (numbering according to Otaka and Osawa 1981; Otaka et al. 1983, respectively). For clarity prefixes Sc and Sp have been added to the numbering of proteins derived from S. cerevisiae and S. pombe, respectively. Ribosomal proteins from these yeasts and from Kluyveromyces marxianus, Rhodotorula glutinis, the slime mold Dictyostelium discoideum and the protozoan Tetrahymena thermophila were checked for antigenic cross-reactivity by the immunoblot technique. Anti-ScL1 bound to the largest ribosomal proteins of all organisms but not with equal strength. A fast migrating protein band from R. glutinis was also reactive. Anti-ScL2 reacted strongly with L2 or analogous proteins derived exclusively from the yeasts. Anti-ScL3 cross-reacted only with one protein band from K. marxianus, whereas anti-SpL1 cross-reacted with L1 or its analogues from the other organisms, but also with proteins of lower molecular weight. In S. cerevisiae, these proteins are located exclusively on the small ribosomal subunit. L2 or analogous ribosomal proteins of all organisms were recognized by anti-SpL2 but additionally the ribosomal protein YL28 of S. cerevisiae and fast migrating proteins of T. thermophila exhibited anti-SpL2 binding.

Animals↗
PubMed1986

Determination of protein by Coomassie dye-binding in agarose gels.

A variation of the Coomassie dye-binding assay for proteins is described. Protein samples were pipetted to the surface of agarose plates in uniformly sized spots and stained with Coomassie Blue G-250. The bound dye was determined by densitometric scanning using double wavelength and flying spot facilities. The response curves were linear in an about 10-fold concentration range with a lower detection limit of 0.5 microgram. No background correction was necessary because unbound dye and most substances known to interfere with other protein assays were removed during the staining and destaining of the agarose gels. Membrane proteins could be analyzed since the samples were applied as solutions in 1% sodium dodecyl sulfate.

Animals↗
PubMed1985

Isolation of yeast ribosomal proteins L3 and L2 for immunological studies.

We report on a rapid method for the isolation and purification of the yeast ribosomal proteins L3 and L2 using a simple instrumentation. Preparative dodecyl sulfate polyacrylamide gel electrophoresis was applied to the separation of cytoplasmatic ribosomal proteins of the large subunit from the yeast Saccharomyces cerevisiae. The polypeptides were removed from gel slices by electrophoretic elution. Subsequent analytical electrophoresis showed groups of proteins in all but two fractions. The latter were further analysed by a two-dimensional gel electrophoresis system which disclosed the purity of two polypeptides. They were identified as L3 and L2. Their molecular masses were 51.5 and 44 kDa as estimated from the gels. A possible application to the isolation of other yeast ribosomal proteins is discussed. An antiserum against the polypeptide L3 was raised in a rabbit. Applying an enzyme-linked immunosorbent assay (ELISA) we were able to determine the relative antibody concentration. Its specificity was demonstrated by immunoblotting.

Antibodies, Fungal↗
PubMed1981

Thiopyronine-sensitized photodynamic effect on RNA synthesis in Saccharomyces cells in vivo.

The effect of photodynamic treatment with thiopyronine and visible light on RNA metabolism in yeast cells was investigated at different times during logarithmic growth. The results show that RNA synthesis in the nucleus of the cells is not directly inhibited photodynamically. In the endoplasmic reticulum of photodynamically treated cells one finds mRNA in about the same relative amounts and quality as in untreated control cells, but the binding of polysomes on membranes in the cytoplasm as the first step of protein synthesis is inhibited for a long time after treatment as well as the synthesis of ribosomes.

Endoplasmic Reticulum↗
PubMed1980

Evaluation of centrifugation parameters for density gradient experiments by means of a programmable pocket calculator.

A calculation program is proposed suitable for programmable pocket calculators (e.g. HP series) to estimate s20,w f omega2 dt values from density gradient centrifugation data. The program can be applied to linear or exponential density gradients prepared from sucrose or glycerol solutions spun in zonal rotors or swinging bucket rotors. A wide solute concentration range and temperature range is accounted for. Constants for empirical density calculation of glycerol and sucrose solutions concentrated in % (w/v) are estimated. Experimental verification of the program was carried out.

Centrifugation, Density Gradient↗
PubMedDate not supplied

Initiation of protein synthesis in yeast: binding of Met-tRNAi.

Conditions for the binding of Met-tRNAi to 40 s ribosomal subunits and to proteins isolated out of the yeast ribosomal KCl wash were investigated. Sucrose density gradient experiments revealed that binding of Met-tRNAi to 40 s ribosomal subunits was catalyzed in a AUG and GTP dependent reaction. Binding of Met-tRNAi to proteins of the ribosomal KCl wash as assayed by the Millipore filter technique was found to be independent of AUG, GTP and 40 s ribosomal subunits. Additions of GTP yielded only slight stimulation, whereas Mg2+ caused dissociation of complexes. It was concluded that these reactions were most likely catalyzed by initiation factor eIF-2 although stimulation by GTP did not occur.

Eukaryotic Initiation Factor-2↗