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Biomedical subjects

H Michel

Publications and source records attributed to H Michel.

At least 163 records · Page 9Linked to original sources
PubMed1993

Orthorhombic crystal form of bacteriorhodopsin nucleated on benzamidine diffracting to 3.6 A resolution.

Freshly formed benzamidine crystals were found to provide a suitable nucleation surface for crystallisation of bacteriorhodopsin. At 20 degrees C and 1% octylglucoside pseudohexagonal needles of bacteriorhodopsin nucleated on the benzamidine surface. At 4 degrees C and reduced detergent concentration (0.5%) a new, better-ordered orthorhombic crystal form of bacteriorhodopsin was formed by heterogeneous nucleation on benzamidine. Polarised absorption spectroscopy and flash photolysis experiments were used to show that the crystalline bacteriorhodopsin is photoactive in both forms. The M-intermediate absorbs maximally at 405nm and formation of M does not disturb the crystal lattice. The plate-shaped crystals diffract to a resolution of 3.6 A along the a and b directions and to 4.2 A in the c direction. The most likely space group of the crystals is C222 (a = 107.5 A, b = 117.0 A, c = 69.5 A). The crystals are built from layers of bacteriorhodopsin molecules that are tilted from the c axis by about 45 degrees. In addition, as a background to the discovery of the new crystal form, the influence of different detergents, additives and precipitants on the formation of pseudohexagonal needles is presented.

Bacteriorhodopsins↗
PubMed1993

[Acute pancreatitis caused by foreign bodies. 2 cases].

We report the cases of two patients with acute pancreatitis caused by the insertion of foreign bodies in the ampulla of Vater: vegetal residues in the first case and beltbuckle in the second case. The acute pancreatitis was revealed by severe epigastric pain, jaundice and a serum amylase increase to more than 9 times the upper limit of normal level. The presence of foreign bodies was detected by endoscopic retrograde cholangiography and pancreatography in the first case and by abdominal X-ray examination in the second case. Extraction of vegetal residues was performed by endoscopic route whereas that of the beltbuckle required surgery. From these 2 cases and 8 others previously reported cases, the clinical features and course of acute pancreatitis caused by foreign bodies have been analysed.

Acute Disease↗
PubMed1993

Synthesis of the Rhodopseudomonas viridis holo-cytochrome c2 in Paracoccus denitrificans.

The gene encoding the Rhodopseudomonas viridis cytochrome c2 (cycA) has been introduced on a broad host range vector into Paracoccus denitrificans, leading to high-level expression of the holo-cytochrome with the heme moiety covalently attached to the apoprotein. The cytochrome was demonstrated to reside in the periplasmic space of the host cell. In contrast to R. viridis, aerobic rather than anaerobic growth conditions led to higher production levels of the holo-cytochrome in P. denitrificans. This heterologous expression system provides a suitable genetic background for the functional expression and mutagenesis of polypeptides involved in bacterial photosynthesis, offering the possibility of detailed structural and functional investigation.

Aerobiosis↗
PubMed1993

Nucleotide sequence of a putative succinate dehydrogenase operon in Thermoplasma acidophilum.

28 amino acids from the N-terminal region of a putative terminal oxidase from the archaebacterium Thermoplasma acidophilum were determined by Edman degradation. On basis of this amino acid sequence a degenerated oligonucleotide was synthesized and used as a radioactive probe for Southern blot analysis of EcoRI digested genomic DNA. A 2.3 kb EcoRI fragment strongly hybridized to the probe and size selected genomic library from genomic DNA was constructed. Several clones scored positive by screening the library with the degenerated oligonucleotide, from which only one clone contained a EcoRI DNA fragment encoding the 28 amino acid sequence determined by protein sequencing. Sequence analysis revealed the presence of three genes in the typical arrangement of an operon. The first gene codes for a protein containing 11 cystein residues in an arrangement typical for Fe/S proteins. Protein sequence comparison revealed significant homologies to the fumarate reductase and succinate dehydrogenase of other bacteria. The two other genes encode small hydrophobic proteins probably serving as membrane anchor for the Thermoplasma acidophilum succinate dehydrogenase.

Amino Acid Sequence↗
PubMed1993

Unexpected similarities of the B800-850 light-harvesting complex from Rhodospirillum molischianum to the B870 light-harvesting complexes from other purple photosynthetic bacteria.

The B800-850 light-harvesting complex (also called LH2) was isolated from photosynthetic membranes of Rhodospirillum molischianum DSM 119 using molecular sieve and ion-exchange chromatography. Its two bacteriochlorophyll a-binding polypeptides (alpha-subunit and beta-subunit) were purified with a reverse-phase HPLC system. The complete amino acid sequences of both subunits have been determined. The alpha- and beta-subunits consist of 56 and 45 amino acids, respectively, corresponding to molecular weights of 5939 and 5133. In contrast to the B800-850 complexes from other photosynthetic bacteria, the native B800-850 complex from Rs. molischianum is most likely an octamer of monomers with a stoichiometry of three bacteriochlorophyll a and 1.5 lycopenes per alpha,beta-subunit. Resonance Raman spectra provide evidence for a 5-coordinated Mg2+ in the BChl, and a carotenoid mainly in the all-trans configuration. A comparison between resonance Raman data from different photosynthetic bacteria indicates that the BChl a-binding site of the B800-850 complex from Rs. molischianum is more similar to the B870 complexes (also called LH1) than to the B800-850 complexes of other photosynthetic bacteria. Sequence similarities especially between the beta-subunits of the B800-850 complex of Rs. molischianum and the B870 and B800-850 complexes of other photosynthetic bacteria agree with this result and provide information on the mode of pigment binding in bacterial antenna complexes.

Amino Acid Sequence↗
PubMed1993

Otoconin-22, the major protein of aragonitic frog otoconia, is a homolog of phospholipase A2.

Otoconia are composites of proteins and inorganic crystals formed in the peripheral portion of the vestibular system of vertebrates. They add mass to the extracellular otoconial membrane, thereby increasing its deflection during linear acceleration. This added mass increases the sensitivity of the underlying sensory maculae. Otoconia provide a promising system to decipher the interaction of protein and mineral during the growth and maintenance of biominerals. We have purified the major protein of the aragonitic otoconia of Xenopus laevis, which we call otoconin-22, and determined its amino acid sequence and carbohydrate composition. The 127 residues are 37% identical to the phospholipase A2 from Crotalus atrox. We propose that otoconin-22 from X. laevis is homologous to phospholipase A2 and has a similar tertiary structure.

Amino Acid Sequence↗
PubMed1993

New crystal form of the photosynthetic reaction centre from Rhodobacter sphaeroides of improved diffraction quality.

Trigonal crystals of photosynthetic reaction centres from the wild-type purple bacterium, Rhodobacter sphaeroides (ATCC 17023), have been grown from potassium phosphate solutions at 18 degrees C. They belong to the space group P3(1/2)21 and have unit cell dimensions of a = b = 141.4 A and c = 187.2 A. The crystals diffract to at least 2.65 A resolution and are suitable for detailed structural studies.

Crystallization↗
PubMed1993

Evidence that serine L223 is involved in the proton transfer pathway to QB in the photosynthetic reaction center of Rhodopseudomonas viridis.

In the reaction center of purple photosynthetic bacteria, the reducing equivalents produced by primary charge separation are exported via an ubiquinone molecule working as a two-electron shuttle. This loosely-bound quinone, called QB, accepts in successive flashes two electrons from the tightly bound primary quinone acceptor QA, along with two protons from the external medium. The surrounding protein plays an important role in stabilizing the semiquinone anion and in providing a pathway for protons from the cytoplasmic phase to QB. Herbicides of the triazine type compete with QB for the binding pocket and their binding is controlled by nearby amino acid residues. We have studied the kinetics of the first and second electron transfer from QA to QB in two herbicide-resistant mutants from Rhodopseudomonas viridis, T1 (ArgL217-->His,Ser L223-->Ala) and MAV5 (Arg L217-->His, Val L220-->Leu), in order to determine whether these residues are involved in proton transfer to the reduced QB. The main effect of the mutant T1 was a drastic (600-fold at pH 7) decrease in the rate of the second electron transfer to QB compared to the wild type. In contrast, the rate of the second electron transfer in the mutant MAV5 was decreased only slightly (10-fold) in the pH range from 7 to 11. We attribute the inhibition of the second electron transfer in the Ser L223-->Ala mutation to an essential role of Ser L223 in the donation of the first proton to the reduced QB.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1993

Molecular structure of a protein-tyrosine/threonine kinase activating p42 mitogen-activated protein (MAP) kinase: MAP kinase kinase.

MAP kinases p42mapk and p44mapk participate in a protein kinase cascade(s) important for signaling in many cell types and contexts. Both MAP kinases are activated in vitro by MAP kinase kinase, a protein-tyrosine and threonine kinase. A MAP kinase kinase cDNA was isolated from a rat kidney library by using peptide sequence data we obtained from MAP kinase kinase isolated from rabbit skeletal muscle. The deduced sequence, containing 393 amino acids (predicted mass, 43.5 kDa), is most similar to byr1 (Bypass of ras1), a yeast protein kinase functioning in the mating pathway induced by pheromones in Schizosaccharomyces pombe. An unusually large insert is present in MAP kinase kinase between domains IX and X and may contribute to protein-protein interactions with MAP kinase. Major (2.7 kilobases) and minor (1.7 kilobases) transcripts are widely expressed in rat tissues and appear to be derived from a single gene.

Amino Acid Sequence↗
PubMed1993

Rat renal, aortic and pulmonary endothelin-I receptors: effects of changes in sodium and water intake.

1. In the present study we investigated, first, the effects of high Na+ intake and, second, the effects of water deprivation on plasma endothelin-1 concentration and urinary endothelin-1 excretion and on endothelin receptors in membranes of renal glomeruli and papillae and of aortic smooth muscle and lung tissue from 32 female Sprague-Dawley rats. 2. After 5 weeks of high Na+ intake (n = 8) urinary Na+ excretion was 10.5 +/- 1.3 compared with 1.6 +/- 0.2 mmol/24 h in controls. Body weight, plasma osmolarity, plasma endothelin-1 concentration (23 +/- 6 versus 28 +/- 3 fmol/ml) and urinary endothelin-1 excretion (6.1 +/- 1.3 versus 4.7 +/- 0.3 pmol/24 h) remained unchanged. 3. The characteristics of endothelin-1 receptors in glomeruli, papillae, aortic smooth muscle and lung tissue from salt-loaded rats were not different from those of controls. 4. After 48 h water deprivation (n = 8) body weight had decreased, whereas packed cell volume and plasma and urine osmolarities had increased compared with controls (n = 8) (P < 0.05). Plasma endothelin-1 concentration (40 +/- 6 versus 21 +/- 2 fmol/ml) was higher (P < 0.01) and urinary endothelin-1 excretion (1.0 +/- 0.2 versus 2.8 +/- 0.3 pmol/24 h) was lower than in controls (P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Characteristics of endothelin receptors and intracellular signalling in porcine inner medullary collecting duct cells.

In porcine kidneys we investigated the characteristics of endothelin (ET) receptors that are present in papillary tissue but not in glomeruli. Therefore, porcine inner medullary collecting duct (IMCD) cells were separated by Percoll density gradient centrifugation after enzymatic and hypotonic treatment of minced papillary tissue. Studies were performed in fresh cell suspensions and in cells in primary culture. Changes in cytosolic free Ca2+ concentration [Ca2+]i were measured by the use of fura-2. Optimum binding of ET-1 was obtained by incubation for 120 min at 37 degrees C, pH 7.0 when maximal protein content was 40 micrograms. Analysis with the LIGAND program showed an average number of binding sites (Bmax) of 26.0 +/- 30.5 fmol/mg protein and dissociation constant (Kd) of 90.5 +/- 28.6 pmol/L for ET-1 and Bmax of 246.9 fmol/mg protein and Kd of 162.5 pmol/L for ET-3. ET-1, 10(-9) to 10(-6) mol/L, dose dependently raised [Ca2+]i four to tenfold, respectively, from a mean basal level of 41 nmol/L. This rise was significantly attenuated by TMB-8 and by verapamil. Preincubation with Ni2+ almost completely prevented the increment in [Ca2+]i. ET-1 slightly suppressed basal and significantly attenuated arginine vasopressin (AVP)-induced cyclic adenosine monophosphate (cAMP) synthesis. Thus, porcine IMCD cells possess a single class of super high affinity ETB receptors (ETB1). ET-1 raises [Ca2+]i through release from intracellular stores, activation of L-type calcium channels and, probably to a larger extent, through stimulation of other channels, eg, T-type calcium channels or unselective cation channels.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Cellular mechanisms of action of a ouabain-like factor in vascular smooth-muscle cells.

Ouabain-like factor (OLF) has been implicated to play an important role in certain forms of hypertension. We isolated OLF from the urine of salt-loaded healthy subjects by stepwise chromatographic procedures. The post-salt fraction (F IV) eluted from Sephadex G-25 was rechromatographed on Sephadex G-10. A late small-molecular-weight fraction F8 inhibited Na-K-ATPase in vitro (OLF activity). The effects of OLF on intracellular Ca2+ concentrations ([Ca2+]i) and pH (pHi) were examined in cultures of vascular smooth-muscle cells using the fluorescent probes fura-2 and 2',7'-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF), respectively. Preincubation with OLF increased basal [Ca2+]i from 87 +/- 6 to 160 +/- 8 nM (p < 0.001) and enhanced arginine vasopressin-stimulated maximal [Ca2+]i (418 +/- 11 vs. 523 +/- 14 nM, p < 0.01). This effect was similar to that of ouabain. OLF also induced a rapid transient increase of [Ca2+]i (82 +/- 9 vs. 253 +/- 23 nM, p < 0.01); [Ca2+]i returned to levels slightly above baseline within approximately 4 min. OLF-stimulated [Ca2+]i was attenuated by verapamil (126 +/- 5 nM, p < 0.01) and was also reduced in Ca(2+)-free medium (104 +/- 9 nM, p < 0.01). As opposed to OLF, ouabain did not exhibit this fast transient effect on [Ca2+]i. Amiloride (10(-3) M) blocked the sustained effect of OLF on [Ca2+]i (77 +/- 11 vs. 86 +/- 12 nM, NS). OLF induced an increase of pHi from 7.09 +/- 0.03 to 7.28 +/- 0.04 (p < 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1993

Structure and expression of chloroplast-localized porphobilinogen deaminase from pea (Pisum sativum L.) isolated by redundant polymerase chain reaction.

Porphobilinogen (PBG) deaminase catalyzes the polymerization of four PBG monopyrrole units into the linear tetrapyrrole hydroxymethylbilane necessary for the formation of chlorophyll and heme in plant cells. Degenerate oligonucleotide primers were designed based on amino acid sequence data (generated by mass spectrometry) for purified PBG deaminase from pea (Pisum sativum L.) chloroplasts. These primers were used in TaqI polymerase-catalyzed polymerase chain reaction (PCR) amplification to produce partial cDNA and nuclear genomic fragments encoding the enzyme. Subsequently, a 1.6-kb cDNA was isolated by screening a cDNA library constructed in lambda gt11 from leaf poly(A)+ RNA with the PCR products. The cDNA encodes an approximately 40-kD polypeptide containing a 46-amino acid NH2-terminal transit peptide and a mature protein of 323 amino acids. The deduced amino acid sequence of the mature pea enzyme is similar to PBG deaminases from other species and contains the conserved arginine and cysteine residues previously implicated in catalysis. Northern blot analysis indicates that the pea gene encoding PBG deaminase is expressed to varying levels in chlorophyll-containing tissues and is subject to light induction.

Amino Acid Sequence↗
PubMed1993

[Failure factors of guided puncture of hepatic masses].

This study was carried out to assess the failure factors of guided hepatic mass puncture in 320 biopsies. Results of the puncture, follow-up or surgery allowed us to divide 320 biopsies into true positives or false negatives. Compared with the group of true positive diagnosis, the group of false negative diagnosis revealed a significantly higher percentage either of lesions less than 2 cm (52.5% versus 34%), of benign tumors (19% versus 5%), or of pseudotumors (24% versus 1.5%). Compared with the group with cytology and histology diagnosis, the group in which only histology established the diagnosis revealed a significantly higher percentage of hepatocellular carcinoma (52.5% versus 34%) and of benign tumors (21% versus 0%). A diameter less than 2 cm, a histology of benign tumors or pseudotumors, are failure factors of guided biopsy. The use of histology is useful for the diagnosis of benign tumors and increases the accuracy of diagnosis for hepatocellular carcinoma.

Biopsy, Needle↗