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K Bock

Publications and source records attributed to K Bock.

At least 73 records · Page 4Linked to original sources
PubMed1994

An NMR spectroscopic and conformational study of 12 pseudo-disaccharides (D-glucopyranosyl-5a-carba-D- and -L-glucopyranoses).

NMR spectroscopic data for 12 pseudo-disaccharides of the general structure: (alpha or beta)-D-glucopyranosyl-(1-->chi)-5a-carba-(D or L)-glucopyranose, representing analogues of laminaribiose (beta-D-Glc p, chi = 3), cellobiose (beta-D-Glc p, chi = 4), and maltose (alpha-D-Glc p, chi = 4) are presented. The assigned NMR chemical shifts together with NOE difference measurements in association with calculations applying the HSEA force field combined with Monte Carlo simulations have been used to assess the conformational preferences of the investigated compounds. The results are correlated with general structural features involved in the interactions between monosaccharide units of oligosaccharides.

Carbohydrate Conformation↗
PubMed1994

Ergot induced peripheral vascular insufficiency, non-interventional treatment.

We report a case of ergotamine tartrate induced severe vasospasm in the renal arteries and the arteries of the lower extremities. Classic features seen on peripheral angiography make the diagnosis. Anticoagulation, thrombolysis, vasodilation, steroids, and prostaglandin inhibitors all have been successfully used to treat symptomatic ergot induced arterial vasospasm. Although balloon angioplasty of ergot induced vasospasm has been described in case reports, ergot vasospasm is a self limited and medically treatable condition that does not require peripheral mechanical intervention, unless the immediate threat of necrosis and gangrene exists.

Adult↗
PubMed1994

Silyl protection in the solid-phase synthesis of N-linked glycopeptides. Preparation of glycosylated fluorogenic substrates for subtilisins.

The trimethylsilyl (TMS) group was used for protection of the hydroxy groups of three disaccharide 1-amino-alditols and of the glycosylamines of glucose, maltotriose and maltoheptose. The per-O-trimethylsilylated derivatives were coupled with N alpha-Fmoc-Asp(Cl)-OPfp 7 to give six glycosylated building blocks for the solid-phase synthesis of N-linked glycopeptides. Building block 8 was used in the synthesis of five internally quenched fluorescent substrates which were studied by enzymatic hydrolysis with savinase, a subtilisin-type enzyme.

Amino Acid Sequence↗
PubMed1994

Susceptibility of glycans to beta-elimination in Fmoc-based O-glycopeptide synthesis.

In order to investigate the possible extent of beta-elimination occurring in Fmoc-based continuous-flow solid-phase glycopeptide synthesis, the influence of the pKb of the base used for N alpha-deprotection has been studied. A glycosylated pentapeptide was synthesized using 50% morpholine, 10% piperidine or 2% DBU, respectively, in DMF for deprotection. The dehydropentapeptide N alpha-Ac-Thr-Thr-delta Aba-Val-Thr-NH2, which would be formed in the case of beta-elimination, was prepared independently and used as a control in HPLC analysis; however, this product was not formed under any of the deprotection conditions applied. Furthermore, a 23 amino acid long glycopeptide from human intestinal mucin was prepared using 2% DBU as a base for Fmoc cleavage, and similarly no beta-elimination was observed. The glycopeptide products were subjected to a prolonged treatment with sodium hydroxide in methanol/water without significant formation of byproducts, and the pure glycopeptides were isolated and characterized by 1H-NMR spectroscopy.

Amino Acid Sequence↗
PubMed1994

HPLC and NMR investigation of the serum amine oxidase catalyzed oxidation of polyamines.

In the presence of amine oxidases polyamines arrest cell proliferation owing to the generation of hydrogen peroxide and amino aldehydes. In this investigation the bovine serum amine oxidase catalyzed oxidation of polyamines has been studied by HPLC analysis of dansylated reaction products with or without NaBH4 reduction and by NMR spectroscopy of the reaction products and 3-aminopropanal was found to be a major reaction product. These findings were further substantiated by analysis of the reaction products by ion-exchange chromatography and by analysis of the products formed by oxidation of polyamines by the cofactor of Cu amine oxidases, 6-hydroxydopa. 3-Aminopropanal is unstable and can give rise to acrolein by beta-elimination.

Acrolein↗
PubMed1994

Albaflavenone, a sesquiterpene ketone with a zizaene skeleton produced by a streptomycete with a new rope morphology.

A novel antibiotic alpha,beta-unsaturated sesquiterpene ketone, albaflavenone with a zizaene skeleton was isolated from a morphologically novel, highly odorous Streptomyces species which was identified with the species group S. albidoflavus, cluster 1. The new compound, partly responsible for the odour, was assigned the structure of 2R',6,7,7-tetramethyl-1S',8R'- tricyclo-[6.2.1.0(1,5)]undec-5-en-4-one based on spectroscopic studies including 2D NMR (COSY, HETCOR, ROESY, NOE-difference) experiments.

Anti-Bacterial Agents↗
PubMed1993

Substrate recognition by amyloglucosidase: evaluation of conformationally biased isomaltosides.

Amyloglucosidase catalyzes the hydrolysis of methyl beta-maltoside (1) 30-50 times more rapidly than methyl alpha-isomaltoside (2). It is established that OH-6', OH-4', and OH-4 which are involved in key polar interactions with the enzyme in the case of isomaltoside. Conformational analyses based on HSEA calculations indicate that the dispositions in space of OH-3 of maltose relative to OH-4' and OH-6' in the preferred conformation for the maltoside (1) is energetically more readily achieved by methyl 6R-C-methyl-alpha-isomaltoside (3), than for its 6-S-isomer (4). A kinetic evaluation of the hydrolysis in fact has shown that the R-compound is more strongly bound by the enzyme (Km = 0.9 mM) than the parent isomaltoside (Km = 24.5 mM), whereas the S-compound has the weakest enzyme binding (Km = 90 mM). Since the kcat values were all within the range 0.85 +/- 0.20 s-1, it is evident that the relative rates of hydrolysis are related to the relative ease for the compounds to achieve an interaction of a hydroxyl group in the aglycon of an alpha-D-glucopyranoside with the enzyme for the formation of the enzyme-substrate complex. The relative rates of hydrolysis of the alpha-glucosides of the 1,3-dihydroxy-trans-decalins, 5 and 6, provide further support for this highly desirable but not necessary recognition for the orientation of the reducing glucose unit in the active site.

Carbohydrate Conformation↗
PubMed1993

The structure of the carbohydrate backbone of the core-lipid-A region of the lipopolysaccharide from Vibrio cholerae strain H11 (non-O1).

Lipopolysaccharide from Vibrio cholerae strain H11 (non-O1) was de-O-acylated, dephosphorylated, reduced, de-N-acylated, N-acetylated, and the products were separated by high-performance anion-exchange chromatography (HPAE). A decasaccharide, 1, was isolated as the major product, representing the core oligosaccharide attached to the reduced GlcN-disaccharide lipid A backbone. Its structure was established by compositional and methylation analyses, and extensive NMR investigations including 1H,1H correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), and nuclear Overhauser enhancement spectroscopy (NOESY), as well as heteronuclear 13C,1H COSY. In another reaction sequence the lipopolysaccharide was hydrolysed with dilute acetic acid and reduced with NaBH4. The resulting core fractions were separated by HPAE giving seven individual octasaccharides differing at the reducing 3-deoxy-D-manno-octulosonic acid (Kdo) residue. A major product, 2, was isolated and investigated by the same methods as described for the decasaccharide 1. The following structures are proposed for compounds 1 and 2: alpha-D-GlcNAcp-(1-7)-[beta-D-Galp-(1-3)-]-alpha-Hepp-(1-2)- alpha-Hepp- (1-3)-[beta-D-Glcp-(1-4)-]- [alpha-D-Glcp-(1-6)-]-alpha-Hepp-(1-5)-R, where R is alpha-Kdop-(2-6)-beta-D-GlcNAcp-(1-6)-D-GlcNAcol in 1 and 4,8-anhydro-Kdool in 2, and Hep is L-glycero-D-manno-heptose. In lipopolysaccharide, the terminal residue of alpha-D-glucosamine possessed a free amino group, as proved by deamination with nitrous acid and the 1H-NMR spectrum of de-O-acylated lipopolysaccharide. The conformational preferences of the terminal core heptasaccharide was assessed by Monte Carlo simulations combined with restrained calculations of side chains based on experimentally determined proton-coupling constants. These calculations, confirmed by NOE data, displayed several long-range interactions, which resulted in a well-defined three-dimensional structure of the core oligosaccharide.

Carbohydrate Conformation↗
PubMed1993

Methods for liquid- and solid-state CP-MAS NMR spectroscopy of untreated tissue biopsies.

We describe a method for NMR analysis of rapidly cooled or frozen biopsies and report its use on rat liver. Since the metabolic state of the biopsies can be expected to reflect the in vivo state, the method may be used as an alternative to the traditional examination of a perchloric acid extract of the biopsy. Perfusion-cooling of rat liver was applied as an efficient means of preserving the metabolic state. This method ensures very rapid cooling without interruption of the oxygen supply to the liver, and the results show that biopsies taken subsequently maintain energy metabolites near in vivo concentrations for at least 40-60 min. High resolution, natural abundance 13C liquid-state NMR spectroscopy could be carried out within this time frame on the untreated biopsy. In addition, the biopsy was frozen for 13C cross polarization-magic angle spinning solid-state spectroscopic examination, which was carried out at -40, -100, and -150 degrees C. The solid-state spectra allowed analysis of the relative glycogen content of the intact liver tissue, which showed good correlation with chemically measured glycogen on the same samples. Furthermore it was observed that the C1-carbon of glycogen in all liver samples splits into two resonances (5.4 ppm apart) in the solid state but not in the liquid state. This suggests that two conformational states of glycogen are populated, with rapid equilibration in the liquid state but no equilibration in the frozen state.

Adenosine Diphosphate↗
PubMed1993

Synthesis and NMR spectroscopic investigation of oligosaccharides containing Kdo and L-glycero-D-manno-heptopyranosyl residues.

The disaccharides O-(sodium 3-deoxy-alpha-D-manno-2-octulopyranosylonate)-(2-->8)-sodium (allyl 3-deoxy-beta-D-manno-2-octulopyranosid)onate (8), O-L-glycero-alpha-D-manno-heptopyranosyl-(1-->7)-sodium (allyl 3-deoxy-beta-D-manno-2-octulopyranosid)onate (12), and O-alpha-D-mannopyranosyl-(1-->7)-sodium (allyl 3-deoxy-beta-D-manno-2-octulopyranosid)onate (21) and the branched trisaccharides O-L-glycero-alpha-D-manno-heptopyranosyl-(1-->7)-[O-(sodium 3-deoxy-alpha- and -beta-D-manno-2-octulopyranosylonate)-(2-->8)]-sodium (allyl 3-deoxy-beta-D-manno-2-octulopyranosid)onate (15 and 16) and O-alpha-D-mannopyranosyl-(1-->7)-[O-(sodium 3-deoxy-alpha-D-manno-2-octulopyranosylonate)-(2-->8)]-sodium (allyl 3-deoxy-beta-D-manno-2-octulopyranosid)onate (24) were prepared. Per-O-acetylated mannopyranosyl or Kdo bromide derivatives were employed for the glycosylation steps under Helferich conditions, whereas the imidate derivative 9 was used for the coupling of the L-glycero-D-manno-heptopyranosyl residues. The oligosaccharides were fully characterized by NMR spectroscopic data. Their structures correspond to an artificial linkage pattern providing a potential cross-reactive epitope for antibodies directed against the inner-core-region of enterobacterial as well as chlamydial lipopolysaccharides.

Carbohydrate Conformation↗
PubMed1993

Word and world order: semantic, phonological, and metrical determinants of serial position.

Animacy, word length, and prosody have all been accorded prominent roles in explanations for word order variations in language use. We examined the sequencing effects of these factors in two types of tasks. In recall tasks designed to simulate language production, we found selective effects of animacy. Animate nouns tended to appear as subjects in transitive sentences, but showed no special affinity for initial position in conjunctions within sentences, but showed no special affinity for initial position in conjunctions within sentences, suggesting a stronger involvement of animacy in grammatical role assignment than in word ordering. Word length had no significant impact: Shorter words did not appear earlier than longer words within sentences or within isolated conjunctions of nouns. Prosody had a weak effect on word order in isolated conjunctions, favoring sequences with alternating rhythm, but only in the absence of an animacy contrast. These results tend to confirm a hypothesized role for conceptual (meaning-based) accessibility in grammatical role assignment and to disconfirm a hypothesized role for lexical (form-based) accessibility in word ordering. In a judgment task, forms with animate nouns early were preferred across all constructions, and forms with short words early were often preferred both in sentences and in conjunctions. The findings suggest a possible asymmetry between comprehension and production in functional accounts of word order variations.

Concept Formation↗
PubMed1992

Active site similarities of glucose dehydrogenase, glucose oxidase, and glucoamylase probed by deoxygenated substrates.

The specificity constants, kcat/KM, were determined for glucose oxidase and glucose dehydrogenase using deoxy-D-glucose derivatives and for glucoamylase using deoxy-D-maltose derivatives as substrates. Transition-state interactions between the substrate intermediates and the enzymes were characterized by the observed kcat/Km values and found to be very similar. The binding energy contributions of individual sugar hydroxyl groups in the enzyme/substrate complexes were calculated using the relationship delta(delta G) = -RT ln [(kcat/KM)deoxy/(kcat/KM)hydroxyl] for the series of analogues. The activity of all three enzymes was found to depend heavily on the 4- and 6-OH groups (4'- and 6'-OH in maltose), where changes in binding energies from 10 to 18 kJ/mol suggested strong hydrogen bonds between the enzymes and these substrate OH groups. The 3-OH (3'-OH in maltose) was involved in weaker interactions, while the 2-OH (2'-OH in maltose) had a very small if any role in transition-state binding. The three enzyme-substrate transition-state interactions were compared using linear free energy relationships (Withers, S. G., & Rupitz, K. (1990) Biochemistry 29, 6405-6409) in which the set of kcat/KM values obtained with substrate analogues for one enzyme is plotted against the corresponding values for a second enzyme. The high linear correlation coefficients (rho) obtained, 0.916, 0.958, and 0.981, indicate significant similarity in transition-state interactions, although the three enzymes lack overall sequence homology.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1992

Elicitors and suppressors of the defense response in tomato cells. Purification and characterization of glycopeptide elicitors and glycan suppressors generated by enzymatic cleavage of yeast invertase.

Cleavage of yeast invertase by alpha-chymotrypsin produced a number of small glycopeptides that were highly active as elicitors of ethylene biosynthesis and phenylalanine ammonia-lyase in suspension-cultured tomato cells. Five of these elicitors were purified and their amino acid sequence determined. They all had sequences corresponding to known sequences of yeast invertase, and all contained an asparagine known to carry a N-linked small high mannose glycan. The most active glycopeptide elicitor induced ethylene biosynthesis and phenylalanine ammonia-lyase half-maximally at a concentration of 5-10 nM. Structure-activity relationships of the peptide part were analyzed by further cleavage of a defined glycopeptide elicitor with various proteolytic enzymes. Removal of the C-terminal phenylalanine enhanced the elicitor activity, whereas removal of N-terminal arginine impaired it. A glycopeptide with the peptide part trimmed to the dipeptide arginine-asparagine was still fully active as elicitor. Glycopeptides with identical amino acid sequences were further separated into fractions differing in the oligosaccharide side chain. A given peptide had high elicitor activity when carrying a glycan with 10-12 mannosyl residues (Man10-12GlcNAc2), a 3-fold lower activity when carrying Man9GlcNAc2 and a 100-fold lower activity when carrying Man8GlcNAc2. The oligosaccharides, released by endo-beta-N-acetylglucosaminidase H from the pure glycopeptide elicitors, acted as suppressors of elicitor-induced ethylene biosynthesis and phenylalanine ammonia-lyase activity. A series of such oligosaccharides in the size range of Man8-13GlcNAc was purified. The structure and composition of the purified oligosaccharides corresponded to the known small high mannose glycans of yeast invertase as verified by 1H NMR spectroscopy at 600 MHz. The highest suppressor activities were obtained with the oligosaccharides containing 10-12 mannosyl residues (Man10-12GlcNAc). The oligosaccharide Man8 GlcNAc was ineffective as a suppressor. Thus, the structural requirements for the free oligosaccharides to act as efficient suppressors were the same as for the oligosaccharide side chains of the glycopeptides for high elicitor activity. We propose that the glycan suppressors bind to the same recognition site as the glycopeptide elicitors without inducing a response.

Amino Acid Sequence↗
PubMed1992

A nuclear magnetic resonance spectroscopic investigation of Kdo-containing oligosaccharides related to the genus-specific epitope of Chlamydia lipopolysaccharides.

The 1H- and 13C-NMR parameters, chemical shifts and coupling constants, for the pentasaccharide of the genus-specific epitope of Chlamydia lipopolysaccharide and related di-, tri-, and tetra-saccharides have been measured and assigned completely using 1D and 2D techniques, and their structures have been confirmed. NOE experiments indicated the preferred conformation of the pentasaccharide and the component oligosaccharides. The 3JH,H demonstrate a change in conformation by rotation of the C-6-C-7 bond of the side chain of the (2----8)-linked Kdo (unit b) in alpha-Kdo-(2----8)-alpha-Kdo-(2----4)-alpha-Kdo-(2----6)-beta-GlcN-(1--- -6)- GlcNol, alpha-Kdo-(2----8)-alpha-Kdo-(2----4)-alpha-Kdo-(2----6)-beta-GlcNAc-(1- ---O)- allyl, and alpha-Kdo-(2----8)-alpha-Kdo-(2----4)-alpha-Kdo-(2----O)-allyl relative to that preferred in alpha-Kdo-(2----4)-alpha-Kdo-(2----6)-beta-GlcNAc-(1----O)-allyl, alpha-Kdo-(2----8)-alpha-Kdo-(2----O)-allyl, alpha-Kdo-(2----4)-alpha-Kdo-(2----O)-allyl, and alpha-Kdo-(2----6)-beta-GlcNAc-(1----O)-allyl, irrespective of the size of the aglycon, e.g., allyl or beta-D-GlcN residues. The conformational results have been substantiated by computer calculations using the HSEA approach.

Antigens, Bacterial↗
PubMed1992

Analysis of conformationally restricted models for the (1----6)-branch of asparagine-linked oligosaccharides by n.m.r.-spectroscopy and HSEA calculation.

The conformational preferences of the trisaccharide, beta-D-GlcpNAc-(1----2)-alpha-D-Manp-(1----6)-beta-D-GlcpOR (1) have been investigated by n.m.r.-spectroscopy and HSEA calculation. The fixed omega-angle bicyclic analogs 2 and 3, models for the gt and gg rotamers, respectively, of 1, were furthermore examined with the same techniques in an attempt to deduce which of the conformations accessible to 1 was recognized and glycosylated by the enzyme GlcNAc-transferase-V, which acts on a component of the (1----6)-arm of glycoproteins. Only the gg bicyclic 3 was found to be reactive with the enzyme and this study concludes, based on conformational analysis, that 1 as well as the natural Asn-linked oligosaccharide are recognized by GlcNAc-transferase-V in only one of the two local minimum energy conformations energetically accessible to these molecules in their gg rotamer.

Asparagine↗
PubMed1992

Regulation of androgen receptor mRNA and protein level by steroid hormones in human mammary cancer cells.

The regulation of the human androgen receptor (AR) by steroid hormones in human mammary cancer cells was investigated using immunocytochemical and ligand binding assays for its protein and Northern blot analyses for the corresponding mRNA. MFM-223 cells contain high levels of ARs and are growth-inhibited by dihydrotestosterone (DHT). The AR protein is down-regulated to 57% of the control by 10 nM DHT after 24 h, and the corresponding mRNA is also reduced. The nonsteroidal antiandrogen hydroxyflutamide had no effect on the AR level, whereas after incubation with 1 microM cyproterone acetate a slight down-regulation was observed. The AR level was restored completely after release from a 7 day treatment with DHT. However, only 60% of the control level was restored, if the cells wer grown in the presence of DHT for 6 weeks. In androgen-pretreated cells the proliferation rate remained decreased even after the withdrawal of DHT. Concomitantly the distinct growth inhibition was lost. Transfection experiments demonstrated a reduced activity of the residual androgen receptor in these pretreated cells. In addition to the AR, EFM-19 cells also contain significant amounts of estrogen and progesterone receptors. EFM-19 cells are not growth inhibited by physiological concentrations of DHT. Autoregulation of AR was also found in this cell line. Additionally, reduced levels of AR protein and mRNA were found in EFM-19 cells after treatment with the synthetic progestin R5020. The maximum effect of R5020 was observed at the high concentration of 1 microM. Estrogen treatment with 10 nM 17 beta-estradiol for 3 days reduced the AR level only by 25%.

Blotting, Northern↗
PubMed1992

From conceptual roles to structural relations: bridging the syntactic cleft.

The distinction between underlying and superficial linguistic structure is a staple of modern cognitive psychology. Despite increasingly diverse conceptions of syntactic relations in linguistic theory, the received view in psycholinguistics has remained one in which the entities assigned to underlying relations may assume different surface relations. The present article examines this view in the context of language production and reviews evidence that the disposition to bind animate entities to the surface subject relation is a basic feature of language use, suggesting that mappings from conceptual categories to syntactic relations form a main support of the bridge from conception to language. Proceeding on this assumption, the article also evaluates competing accounts of the mapping process in production. The results argue against syntactic relation-changing operations, but favor a division between meaning- and form-related mechanisms.

Concept Formation↗