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J Meyer

Publications and source records attributed to J Meyer.

At least 433 records · Page 24Linked to original sources
PubMed1995

Identification and characterization of the dystrophin anchoring site on beta-dystroglycan.

Dystrophin, the product of the Duchenne muscular dystrophy gene, is tightly associated with the sarcolemmal membrane to a large glycoprotein complex. One function of the dystrophin-glycoprotein complex is to link the cytoskeleton to the extracellular matrix in skeletal muscle. However, the molecular interactions of dystrophin with the membrane components of the dystrophin-glycoprotein complex are still elusive. Here, we demonstrate and characterize a specific interaction between beta-dystroglycan and dystrophin. We show that skeletal muscle and brain dystrophin as well as brain dystrophin isoforms specifically bind to beta-dystroglycan. To localize and characterize the dystrophin and beta-dystroglycan interaction domains, we reconstituted the interaction in vitro using dystrophin fusion proteins and in vitro translated beta-dystroglycan. We demonstrated that the 15 C-terminal amino acids of beta-dystroglycan constituted a unique binding site for the second half of the hinge 4 and the cysteine-rich domain of dystrophin (amino acids 3054-3271). This dystrophin binding site is located in a proline-rich environment of beta-dystroglycan within amino acids 880-895. The identification of the interaction sites in dystrophin and beta-dystroglycan provides further insight into the structure and the molecular organization of the dystrophin-glycoprotein complex at the sarcolemma membrane and will be helpful for studying the pathogenesis of Duchenne muscular dystrophy.

Amino Acid Sequence↗
PubMed1995

Direct measurement of lipid peroxidation in submitochondrial particles.

The susceptibility of the polyunsaturated fatty acid parinaric acid (cis-PnA) to peroxidative damage with concomitant loss of its fluorescent character can be used to detect lipid peroxidation in a direct and sensitive way. The procedure, originally developed to measure peroxidation in lipid vesicles and erythrocyte membranes, has been adapted for the study of submitochondrial particles. Optimal conditions for the concentrations of cis-PnA (0.8 mol %), mitochondrial membrane (100 microM membrane phospholipid), and the radical generating system (50 microM NADH and 10 microM:1 mM Fe(III)-ADP) were established. In the absence of peroxidation inducing compounds, a stable fluorescent signal can be detected. Upon addition of NAD(P)H and ADP-Fe(III), lipid peroxidation starts, and the observed fluorescence decrease is a measure of peroxidation. Both NADH and NADPH were able to induce lipid peroxidation in submitochondrial particles in the presence of an iron chelate. The use of NADH resulted in higher rates of peroxidation compared with NADPH at the same concentration. Whereas the rate of NADH-induced lipid peroxidation was maximal at very low NADH concentrations (2.5 microM) and decreased when the concentration became higher, the NADPH-induced lipid peroxidation reaches saturation at 100 microM. NADH-induced lipid peroxidation in submitochondrial particles from different rat tissues (heart, skeletal muscle, and liver) resulted in a clear difference in peroxidation rates. The highest rates were observed in heart submitochondrial particles, while the lowest rates were obtained in submitochondrial particles derived from liver. Skeletal muscle submitochondrial particles showed intermediate rates of lipid peroxidation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Diphosphate↗
PubMed1995

Characterization of a mutated rubredoxin with a cysteine ligand of the iron replaced by serine.

The active site of rubredoxins consists of a single iron tetrahedrally coordinated to four cysteinate sulfurs. One of the iron ligands, cysteine 42, has been mutated into serine in Clostridium pasteurianum rubredoxin. This mutation resulted in a shift to higher energy of the 320-800 nm region of the UV-visible absorption spectrum. Resonance Raman spectra showed that the nu 1 breathing mode of the iron chromophore was upshifted as a result of the C42S mutation. The spectral pattern, however, was not largely disturbed by the mutation. The EPR spectra of both the wild type and the C42S mutated protein displayed the characteristic features, at g = 4.3 and g = 9.5, of the "3/2" and "1/2" Kramers' doublets, respectively, of a S = 5/2 multiplet. These combined data afford strong evidence that in the C42S mutated rubredoxin serine has replaced cysteine 42 as a ligand of the iron, while maintaining the tetrahedral coordination of the metal. The most spectacular effect of the C42S mutation was a ca. 200 mV downshift of the redox potential of rubredoxin.

Base Sequence↗
PubMed1995

Observation of holoprotein molecular ions of several ferredoxins by electrospray-ionization-mass spectrometry.

Several ferredoxins containing [4Fe-4S] or [2Fe-2S] active sites have been analyzed by electrospray-ionization-mass spectrometry. For these acidic proteins, low pH conditions must be implemented in order to ensure strong signals in positive-ionization mode. Under such conditions the iron-sulfur active sites were lost in most cases. In contrast, the holoproteins were preserved under negative-ionization mode conditions: they were weakly but sufficiently ionized and information about their cofactor content could be obtained. The experimental conditions set up here should provide a useful basis for the detailed characterization of more complex iron-sulfur proteins.

Ferredoxins↗
PubMed1995

Synergistic interaction of adenylate cyclase activators and nitric oxide donor SIN-1 on platelet cyclic AMP.

The molecular mechanism of the synergistic platelet inhibition by activators of adenylate cyclase and guanylate cyclase in human platelets was investigated. The adenylate cyclase activators iloprost and prostaglandin E1 and the guanylate cyclase activator 3-morpholino-syndnonimine (SIN-1) dose-dependently inhibited thrombin-induced aggregation of washed human platelets. Furthermore, SIN-1 at a concentration inhibiting platelet aggregation by only 10% shifted the IC50 values of iloprost and prostaglandin E1 by one order of magnitude to the left, indicating a synergistic action of adenylate cyclase and guanylate cyclase activators. Iloprost and prostaglandin E1 dose-dependently elevated platelet cAMP without a significant influence on cGMP. In contrast, the platelet cGMP level was dose-dependently elevated by SIN-1. In addiiton, SIN-1 markedly increased cAMP level induced by low concentrations of adenylate cyclase activators (0.1-0.3 nM iloprost or 10-150 nM prostaglandin E1). In contrast, the rise in cAMP induced by higher adenylate cyclase activator concentrations (3 nM iloprost or 30 microM prostaglandin E1) was significantly reduced in the presence of SIN-1. The same biphasic mode of action of SIN-1 was observed with forskolin, an adenylate cyclase stimulator acting receptor independently, indicating a prostacyclin-receptor independent mechanism. The cAMP elevating effect of SIN-1 in the presence of low prostanoid concentrations was completely abolished by piroximone, a selective inhibitor of phosphodiesterase type III. Therefore, the inhibition of phosphodiesterase III by cGMP seems to be the mechanism for the elevation of cAMP levels by SIN-1 in the presence of low concentration of adenylate cyclase activators in human platelets.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗
PubMed1995

Electrospray-ionization mass spectrometry of molecular variants of a [2Fe-2S] ferredoxin.

The [2Fe-2S] ferredoxin from Clostridium pasteurianum is a homodimeric protein of which each subunit contains one [2Fe-2S] cluster. In previous investigations, the five cysteine residues in positions 11, 14, 24, 56 and 60 had been mutated into serine or alanine. The wild type ferredoxin and several of its molecular variants have now been analyzed by electrospray-ionization mass spectrometry. In the negative-ion detection mode, depending on the infusion solvent used, molecular peaks attributable to the apoprotein, to the monomeric holoprotein, and to the dimeric holoprotein were detected in all cases. The data confirmed the presence of the expected mutations, showed that all of these proteins contain one [2Fe-2S] cluster per subunit, and indicated that the dimeric structure of these ferredoxins could be retained in the conditions of the electrospray ionization. This investigation establishes the power of electrospray-ionization mass spectrometry for the analysis of oligomeric proteins containing labile metal clusters.

Alanine↗
PubMed1995

SH3 domain-mediated interaction of dystroglycan and Grb2.

Dystroglycan is a novel laminin receptor that links the extracellular matrix and sarcolemma in skeletal muscle. The dystroglycan complex containing alpha- and beta-dystroglycan also serves as an agrin receptor in muscle, where it may regulate agrin-induced acetylcholine receptor clustering at the neuromuscular junction. beta-Dystroglycan has now been expressed in vitro and shown to directly interact with Grb2, an adapter protein involved in signal transduction and cytoskeletal organization. Protein binding assays with two Grb2 mutants, Grb2/P49L and Grb2/G203R, which correspond to the loss-of-function mutants in the Caenorhabditis elegans sem-5, demonstrated that the dystroglycan-Grb2 association is through beta-dystroglycan C-terminal proline-rich domains and Grb2 Src homology 3 domains. Affinity chromatography has also shown endogenous skeletal muscle Grb2 interacts with beta-dystroglycan. Immunoprecipitation experiments have demonstrated that Grb2 associates with alpha/beta-dystroglycan in vivo in both skeletal muscle and brain. The specific dystroglycan-Grb2 interaction may play an important role in extracellular matrix-mediated signal transduction and/or cytoskeleton organization in skeletal muscle that may be essential for muscle cell viability.

Adaptor Proteins, Signal Transducing↗
PubMed1995

Cardiac output in single-lead VDD pacing versus rate-matched VVIR pacing.

The importance of atrioventricular synchronous pacing compared with single-chamber rate-responsive pacing is still under discussion, especially for low-intensity workload representing daily life activities. We evaluated hemodynamics in single-lead VDD pacing versus VVIR pacing in 11 patients (8 men and 3 women, aged 58.6 +/- 13.8 years) with normal left ventricular function and a previously implanted single-lead VDDR pacemaker. A low-intensity steady-state treadmill test at 1 to 2.5 mph with a gradient of 2% to 4% was performed. Cardiac output was determined using a standard carbon dioxide rebreathing technique. Initially, the VDD mode was programmed, and after 5 minutes of exercise, cardiac output was measured in steady-state conditions. The pacemaker was then reprogrammed to the VVI mode at a rate 5 to 10 beats above the maximal atrial tracking rate to simulate rate-matched VVIR pacing (VVIRm). After 5 additional minutes of steady-state exercise, cardiac output was measured again. The maximal atrial rate in the VDD mode was 119 +/- 19 beats/min versus a programmed rate of 129 +/- 18 beats/min in the VVIRm mode. VDD pacing resulted in a significantly higher cardiac output than VVIRm pacing (10.6 +/- 1.9 vs 9.2 +/- 1.4 L/min; p < 0.002), with a mean difference of 1.6 +/- 1.2 L/min between the 2 modes. In the VDD mode, stroke volume (90.7 +/- 20.1 vs 71.6 +/- 13.0 ml; p < 0.001) and maximal oxygen uptake (1,183 +/- 264 vs 1,076 +/- 289 ml/min, p < 0.01) were also higher than in VVIRm.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

1,25(OH)2D3-dependent regulation of calbindin-D28k mRNA requires ongoing protein synthesis in chick duodenal organ culture.

Organ culture of 19-day-old chick embryo duodena was utilized to evaluate the mechanism of 1,25-dihydroxyvitamin D3 (1,25(OH)2D3)-dependent calbindin-D28k (CaBP) expression. Duodenal CaBP and 1,25(OH)2D3 receptor (VDR) expression were assessed by Western blot analysis, while CaBP and VDR mRNA levels were determined by Northern blot analysis. In untreated duodena, both VDR protein and mRNA were present, while CaBP protein and mRNA were undetectable. Treatment of cultured duodena with 25 nM 1,25(OH)2D3 resulted in detectable CaBP mRNA after 4 h which continued to increase during a 24 h time period. Under these conditions, localization of [3H-1 beta]1 alpha,25(OH)2D3 in duodenal chromatin is rapid (< or = 30 min). Thus, the delayed accumulation of detectable CaBP mRNA cannot be explained by slow nuclear binding of 1,25(OH)2D3. The inclusion of 1.6 microM actinomycin D in the organ culture partially inhibited the 1,25(OH)2D3-regulated increase in CaBP mRNA, which implies that there is a transcriptional component involved in the increased CaBP mRNA levels. Similarly, quantitative polymerase chain reaction studies allowed the detection of CaBP pre-mRNA and mRNA sequences 1 h after hormone treatment, suggesting that CaBP gene transcription is initiated rapidly. Treatment of cultures with 36 microM cycloheximide 1 h prior to 1,25(OH)2D3 addition resulted in superinduction of VDR mRNA levels but sharply reduced CaBP steady-state mRNA levels. This dramatic reduction in CaBP mRNA reveals that 1,25(OH)2D3-mediated CaBP expression is dependent on ongoing protein synthesis. Thus, we propose that a labile auxiliary protein or other cofactor, which may or may not be 1,25(OH)2D3-dependent, is necessary for 1,25(OH)2D3-mediated CaBP gene transcription in chick duodena.

Animals↗
PubMed1995

Sequence of a 10.5 kbp fragment of Clostridium pasteurianum genomic DNA encompassing the hydrogenase I gene and two spore germination genes.

The gene encoding hydrogenase I from Clostridium pasteurianum had previously been cloned and sequenced as part of a 2.3 kbp Sau3A genomic DNA fragment. The analysis of the regions surrounding the hydrogenase gene has been extended by cloning and sequencing two EcoRI fragments, both partially overlapping the 2.3 kbp Sau3A fragment. An uninterrupted genomic sequence of 10.5 kbp has thus been obtained, including 6.7 kbp upstream of the hydrogenase gene, and 2 kbp downstream. The hydrogenase gene is separated by 473 bp from the next open reading frame on its 5' side and by 366 bp from the next open reading frame on its 3' side. It is preceded by putative promoter regions, and followed by a strong transcription termination signal. Therefore the hydrogenase I gene from C. pasteurianum probably belongs to a monocistronic operon. This is consistent with previous biochemical evidence showing that the enzyme is monomeric. The sequence data also show that in the case of this Fe-hydrogenase, in contrast to the NiFe-hydrogenases, there are no accessory genes cotranscribed with, or located near, the structural genes. The sequenced region contains six open reading frames in addition to the hydrogenase gene. One of these probably encodes phosphatidylserine decarboxylase, and two others are homologous to two of the three genes of the gerA and gerB loci (spore germination) from Bacillus subtilis. These are the first spore germination gene sequences obtained from a bacterium of the genus Clostridium.

Journal Article↗
PubMed1995

Symptom prevalence and worry about high voltage transmission lines.

Few studies have documented public perceptions of environmental health risks from exposure to overhead transmission lines. In particular, little information has been provided on the impact of worry on symptom prevalence in residents living adjacent to high voltage transmission lines. The current study assessed symptom prevalence and worry in 152 Orange County female residents living either adjacent to overhead transmission lines or one block away. Forty-five percent of the respondents were either very worried or somewhat worried about the transmission lines and 55% were slightly worried or not worried at all. Results indicated that for those who did not live on the easement level of worry did not affect the prevalence of health problems. For those who lived on the easement, the most worried respondents were more likely to report health problems. Disclosure of health problems may depend more on individuals' level of worry about rather than proximity to overhead transmission lines. Possible limitations of this study include personality variables such as hypochondriasis which were not assessed, recall bias, and social desirability. The homogeneity of subjects may also limit the generalizability of findings.

Adult↗
PubMed1995

Diminished inhibition of adhesion molecule expression in prostacyclin receptor desensitized human platelets.

Long-term exposure of platelets to prostacyclin or iloprost (100nM, 3hr) results in receptor desensitization measured as decrease in 3H-iloprost binding sites by 47 +/- 14%. Desensitized platelets respond with an increased adhesion to endothelial cells. The mechanism of increased adhesiveness was studied by measuring the expression of the adhesion molecule CD62p (p-selectin; GMP140) on washed human platelets by flowcytometry. In thrombin stimulated platelets CD62p expression was dose-dependently reduced by iloprost. In receptor desensitized platelets IC50 for iloprost inhibition of thrombin-induced CD62p expression increased from 0.48 +/- 0.10 to 2.4 +/- 0.7 nM.

Analysis of Variance↗
PubMed1995

Saruplase in Myocardial Infarction.

Saruplase is an unglucosylated single-chain recombinant urokinase-type plasminogen activator. Dose finding studies in patients with acute myocardial infarction indicated that a dose of 80 mg of saruplase, given as a bolus of 20 mg and iv infusion of 60 mg in one hour, led to excellent patency figures.Saruplase is most effective when combined with a bolus of 5000 IU heparin followed by an iv heparin infusion for at least 24 hours.When saruplase is compared to other thrombolytic agents (streptokinase, alteplase, urokinase), it becomes apparent that its profile is excellent. Early patency rates are at least comparable to alteplase. Further reocclusion rates of saruplase after one day are lower than those of streptokinase and alteplase. Patency rates 24-72 hours after start of medication are comparable between saruplase and urokinase.The large database in over 6000 patients shows that saruplase, in comparison to the other thrombolytic agents, is safe. Its bleeding complication rate is significantly lower than streptokinase, and a trend to lower in-hospital mortality is observed when compared to urokinase.Summarizing, when comparing to the presently available thrombolytic agents, saruplase is a fast acting, effective and safe thrombolytic agent.

Journal Article↗
PubMed1995

Improvement of coronary morphology and blood flow after stenting. Assessment by intravascular ultrasound and intracoronary Doppler.

Intravascular ultrasound (IVUS) and intracoronary Doppler (ICD) were performed in eight patients (54.3 +/- 6.5 years, 6 male) immediately after PTCA and after stenting. ICD was also performed before PTCA. After PTCA, IVUS has demonstrated intimal rupture in all patients. After stenting, IVUS revealed wall wrapping of the intimal flap with a free lumen in all patients. The lumen diameter was 2.42 +/- 0.55 mm after PTCA and was 2.74 +/- 0.49 mm after stenting (p < 0.001). The cross-sectional area increased from 4.70 +/- 1.99 mm2 post-PTCA to 6.40 +/- 2.15 mm2 post-stent (p < 0.005). Coronary flow velocity reserve, calculated by the ratio of mean flow velocity at rest and after intracoronary papaverine administration, increased from 2.05 +/- 1.01 to 2.99 +/- 1.14 after PTCA (p = 0.015); and increased to 4.51 +/- 1.33 after stenting (p < 0.001). The morphological data derived from IVUS correlated with the functional information obtained with ICD. In addition to its established role in bail out situations, stent implantation may be considered when a suboptimal morphological and functional result has been demonstrated.

Angioplasty, Balloon, Coronary↗