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K S Boos

Publications and source records attributed to K S Boos.

36 records · Page 2Linked to original sources
PubMed1988

On-line sample processing and analysis of diol compounds in biological fluids.

We developed a coupled dual column system with an optional post-column derivatization for on-line sample processing, trace enrichment and analysis of aromatic 1,2-diol and aliphatic cis-diol biomolecules (e.g. catecholamines, ribonucleosides). The fully automated high-performance liquid chromatography analyzer tolerates the direct injection of proteinaceous fluids by use of a unique bonded-phase precolumn material which allows the simultaneous performance of covalent affinity and size-exclusion chromatography.

Catecholamines↗
PubMed1988

A minimum structured adenine nucleotide for ADP/ATP transport in mitochondria.

An ADP analogue, i.e. 3H- or 14C-labeled 5'-diphosphate of 6-(beta-D-ribofuranosyl-methyl)-4-pyrimidinamine, was synthesized, the structure of which was deduced from structure-activity studies on the substrate specificity of the nucleotide-binding center of the inner mitochondrial membrane integrated ADP/ATP carrier protein. Bearing only the minimal but substrate-essential recognition structures, minimum structured ADP (msADP) is bound to the cytosol-facing active center and transported by the highly specific carrier system across the inner mitochondrial membrane.

Adenosine Diphosphate↗
PubMed1986

Direct clean-up and analysis of ribonucleosides in physiological fluids.

We describe the group-selective separation and quantification of unmodified, modified and hypermodified ribonucleosides in physiological fluids (urine, serum) by on-line multidimensional high-performance affinity chromatography (HPAC)-reversed-phase liquid chromatography (RPLC). The excretion levels and patterns of ribonucleosides such as N1-methyladenosine, N1-methylinosine, N2-methylguanosine, N2-dimethylguanosine, N6-carbamoylthreonyladenosine and 2-pyridone-5-carboxamido-N-ribofuranoside were determined in urines from a control group and from patients with different diseases. The HPAC-RPLC method applied represents a powerful tool, e.g. as a non-invasive screening test, a method to investigate disorders in ribonucleoside and/or RNA metabolism, a method for drug monitoring during nucleoside chemotherapy, and a method to study renal ribonucleoside reutilization.

Adolescent↗
PubMed1984

Development of a chromatographic method for the quantitative determination of minor ribonucleosides in physiological fluids. Characterization and quantitative determination of minor ribonucleosides in physiological fluids, Part I.

We describe an on-line multi-column high performance liquid chromatographic method for the selective clean-up and analysis of major and minor ribonucleosides in physiological fluids. Quantitative data obtained for the determination of some methylated ribonucleosides in human urines are compared with those obtained with the traditional off-line method. The on-line technique developed in our laboratory is distinguished from the off-line method by the following features: Sample clean-up and analysis of the target-compounds can easily be automatized, Total time of analysis, for example of urinary ribonucleosides, is decreased to 35 minutes, Laborious and error-prone evaporation and redissolution steps are avoided, Reliability of the overall analytical system can be controlled with ease, Small sample-volumes can be applied directly, Sensitive samples can be processed very rapidly under mild conditions, Results obtained with the on-line and off-line-techniques compare well.

Boronic Acids↗
PubMed1983

On-line high-performance liquid affinity chromatography-high-performance liquid chromatography analysis of monomeric ribonucleoside compounds in biological fluids.

We describe an on-line multi-dimensional column chromatography system. It consists of a high-performance liquid affinity chromatography perfractionation column for cis-diol compounds and a series-connected reversed-phase high-performance liquid chromatography system for the analysis of ribonucleosides. This on-line procedure allows the rapid and direct analysis of methylated ribonucleic acid catabolites in biological fluids (serum, urine) and therefore might be useful in pathobiochemistry.

Chemical Phenomena↗
PubMed1983

A model for metalloprotein-catalysed ADP, ATP transport in mitochondria.

We propose a hypothetical model for the transmembrane exchange reaction catalysed by the mitochondrial adenine nucleotide carrier protein, which basically consists of an alternating reorientation of a transitory carrier-metal-nucleotide complex. The key features of the model are: the participation of an intrinsic divalent metal ion in the course of transport catalysis; the different stability constants of protonated and deprotonated nucleotide-metal complexes; the exposure and retraction of strategic arginyl residues; the alternating reorientation of the active center involving a change from the cytosolic conformation (Cc) to the matrix conformation (Cm).

Animals↗
PubMed1983

Inhibition study of ADP,ATP transport in mitochondria with trinitrophenyl-modified substrates.

The ADP,ATP carrier of rat liver mitochondria is specifically inhibited by Meisenheimer-type trinitrophenyl (TNP) derivatives of ADP and ATP. Due to a systematic inhibition study we could show that the TNP-moiety itself, even in the 2', 3'-O-cyclic Meisenheimer complex, revealed no inhibition of mitochondrial ADP,ATP transport. Nucleosidic TNP-compounds are weak inhibitors. Introduction of a phosphate group at the 5'-position, however, enhances the inhibitory power markedly. Our findings point to a synergistic effect of the 5'-phosphate chain and the TNP-moiety. Irreversible inhibition by TNP-nucleotides can be ruled out to the fact that the inhibited ADP,ATP-transport system is fully reactivated by addition of albumin.

Adenosine Diphosphate↗
PubMed1982

ADP and ATP transport in mitochondria. Evidence for a metal-ion involved in transport catalysis by use of metallochromic indicators.

This study introduces a new class of active-site directed probes with respect to ADP and ATP transport catalysis in rat liver mitochondria. The anionic monoazo dyes, e.g., p-(2-hydroxy-1-naphthylazo)naphthol-sulfonic acid, are competitive inhibitors of carrier-mediated ADP uptake (Ki 20-30 microM). The azo dyes also can displace the same amount of carrier-specific bound ADP as does carboxyatractyloside. Two essential substructures could be derived from a structure-activity study. Firstly, a sulfonic acid group in the para position relative to the azo bridge which becomes neutralized upon binding by a specifically located positive charge of the carrier protein. This electrostatic binding component, which presumably is represented by a strategic arginyl residue, seems to be essential for substrate binding as well as inhibitor binding. The second structural requirement for effective inhibition was found to be the o-hydroxy or o,o'-dihydroxyazo system, which is known to form stable complexes with metal ions by chelation. Experiments on prevention and reversal of dye-mediated inhibition revealed that the metal-chelating properties are responsible for the effects observed. In addition, using bovine serum albumin or the synthetic polymer Kollidone, inhibition could be prevented as well as abolished. It is postulated that a metal ion, possibly Mg2+, which is bound to the carrier protein plays an essential role for transport catalysis. The metal ion is assumed to form a functional ternary complex, i.e., a metal bridge complex between the carrier protein and its substrate.

Adenosine Diphosphate↗
PubMed1980

Adenosine di- and triphosphate transport in mitochondria. Role of the amidine region for substrate binding and transport.

A variety of base-modified nucleotide analogues was prepared and characterized as their alpha-32P- or U-14C-labeled compounds. Carrier-linked nucleotide binding and carrier-catalyzed exchange across the inner membrane of rat liver mitochondria were measured by using an inhibitor (atractyloside) stop method. Kinetic data of carrier-specific bound analogues were evaluated from Dixon plots and indicate that these analogues are competitive inhibitors for mitochondrial [14C]ADP uptake. Km and Vmax values for carrier-mediated uptake of nucleotide analogues were calculated from Lineweaver-Burk plots. By means of the analogues, a systematic mapping of the essential chemical and steric interactions between the transporter protein and the heterocycle of its substrate in the course of the binding as well as transfer step was achieved. Prerequisites for carrier-specific binding (recognition) are (A) an anti- or syn-positioned beta-glycosyl-linked heterocycle, (B) a nitrogen ring atom in position 7 for syn-structured analogues, and (C) an electron-rich region at the N(1) position, i.e., a permanent dipole moment oriented toward N(1) for anti-structured analogues. Additional requirements for subsequent transport catalysis are (A) a non-fixed anti-positioned base moiety with a beta-glycosyl torsion angle of about -20 degrees, (B) a C(6)-positioned amino group, and (C) an unsubstituted C(2) atom. The complementary binding site at the carrier protein to the N(1)-C(6)(-NH2) amidine region is proposed to be represented by two juxtaposed and invariant bonding points, i.e., an asparagine or glutamine residue.

Adenosine Diphosphate↗
PubMed1975

Properties of the ribose-ring-opened adenine nucleotide, 2,2'(1-(9-adenyl)-1'-(tri-,diphosphoryl-oxymethyl))-dihydroxydiethylether in mitochondrial adenine-nucleotide translocation.

(Adenine-14C) or (gamma-32P)-labelled 2,2'[1-(9-adenyl)-1'-(tri-, diphosphoryl-oxymethyl]-dihydroxydiethylether (rroANP) was obtained from ANP by cleavage of the C-2--C-3' bond by sodium periodate oxidation and subsequent borohydride reduction. Binding of rroANP to rat liver mitochondria revealed carrier-linked (atractyloside-sensitive) and unspecific (atractyloside-insensitive) binding but no transfer across the inner mitochondrial membrane. Kinetic data indicate rroANP as a competitive inhibitor for ANP uptake with Ki = 9.3 X 10(-5) M. Experimental rroANP confirmed that an intact adenine base and three anio.nic charges of the phosphate chain are essential for the recognition between ANP-carrier and nucleotide but insufficient for the induction of a transmembrane ANP exchange. In addition mobilisation of the carrier-nucleotide complex requires an intact ribofuranoside ring system.

Adenine Nucleotides↗
PubMedDate not supplied

Conformationally restricted adenine nucleotide analogs in mitochondrial adenine nucleotide transport.

The conformationally restricted adenine nucleotide analogs 8,3'anhydro-8-oxy-9-(beta-D-xylofuranosyl)adenine-5'-O-tri(di)-phosphate (I), and 8,2'-anhydro-8-oxy-9-(beta-D-arabinofuranosyl) adenine-5'-O-tri(di)-phosphate (II), were prepared chemically as their alpha-32P-labelled compounds and compared with syn-structured 8-bromo AT(D)P in mitochondrial adenine nucleotide translocation. The experimental findings demonstrate that the heterocycle-ribose orientation affects the carrier mediated adenine nucleotide transport very strongly, i.e. a non fixed adenine heterocycle in the anti region is prerequisite for the bound nucleotide to induce the transfer action of the adenine nucleotide carrier.

Adenine Nucleotides↗