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Biomedical subjects

N Hampp

Publications and source records attributed to N Hampp.

10 recordsLinked to original sources
PubMed2001

Enzymatic cross-linking of purple membranes catalyzed by bacterial transglutaminase.

It was found that bacterial transglutaminase (TGase) facilitates selective cross-linking of bacteriorhodopsin (BR) in purple membrane (PM) form under mild conditions. Fluorescent probes were used to detect that the membrane protein BR may act as a glutamine donor as well as a lysine donor for TGase. The binding sites were determined to be Gln-3 as the reactive glutamine, and Lys-129 is the corresponding lysine residue. Upon incubation of PM with TGase, cross-linking of PM patches can be achieved without an additional spacer molecule. To our knowledge, this is the first time that an intermembrane cross-linking of membrane-bound proteins is reported. Furthermore, this finding may provide the ability to achieve covalent linkage of complete purple membrane patches to synthetic polymers.

Bacteriorhodopsins↗
PubMed1996

Poly(methylene blue)-modified thick-film gold electrodes for the electrocatalytic oxidation of NADH and their application in glucose biosensors.

Electropolymerization of the phenothiazine derivative methylene blue (MB) on screen-printed, thick-film gold electrodes leads to electrocatalytically active and conducting layers of poly(methylene blue) (PMB) in intimate and stable contact with the electrode surface. The catalytic properties of the PMB films allow anodic oxidation of NADH at potentials as low as +200 mV vs. the saturated calomel electrode (SCE) reducing interferences from cooxidizable species as well as minimizing electrode fouling by enabling a simultaneous two-electron transfer mechanism. Dehydrogenase-based biosensors employing PMB-modified thick-film electrodes are obtained either by entrapment of the enzyme into the PMB layer itself or by laminating an enzyme membrane made of an aqueous poly(vinylacetate) dispersion over the PMB-modified electrode. Both methods are used to fabricate glucose biosensors which can be operated at low overpotentials, i.e. +200 mV vs. SCE.

Biosensing Techniques↗
PubMed1994

Potentiometric thick-film sensor for the determination of the neurotransmitter acetylcholine.

A potentiometric thick-film biosensor for the determination of the neurotransmitter acetylcholine is presented. The selective membranes consist of two layers. The first is a choline- and H(+)-selective membrane made from a poly(vinyl chloride)-poly(vinyl acetate) copolymer which is plasticized with 2-nitrophenyl octyl ether. It contains 1% m/m of the ion-pair choline phosphotungstate as an electroactive compound. Both products of the enzymic cleavage, choline and H+, are detected. The second layer is placed above the first and is made from a poly(vinyl acetate)-polyethylene dispersion polymer. It is loaded with 2% m/m of acetylcholinesterase isolated from electric eel. The linear region of the response curve extends from 2 x 10(-5) to 0.01 mol dm-3 in buffer solution and undiluted horse serum. The detection limit is 5 x 10(-6) mol dm-3 and the response time is about 4 min.

Acetylcholine↗
PubMed1993

Photochemical conversion of the O-intermediate to 9-cis-retinal-containing products in bacteriorhodopsin films.

The photochemical activity of the O-state was investigated in bacteriorhodopsin (BR) films containing wildtype BR at pH 6.5 in the presence of glycerol. The formation of a photoproduct of O with an absorption maximum at 490 nm and 9-cis-retinal configuration was found. This 490-nm product was named P and shows a slow thermal reaction into a compound with a maximal absorption at 380 nm which was named Q and contains free 9-cis-retinal in the proteins binding site. The photoproducts of O, i.e., P and Q, are very similar, or even identical, to those previously observed in blue membranes. Common to the O-state and blue membrane forms of bacteriorhodopsin is a protonated aspartic acid 85, and we suggest that it is the reduced negative charge around the Schiff base which is responsible for the 9-cis photoisomerization. The release of a proton from aspartic acid 85 is linked to the conversion of the O-state back to the initial state of BR. Therefore the conditions of low proton mobility in BR films containing glycerol favor the accumulation of the O-state. For optical and holographic applications such BR films are very attractive. It is possible to create photoproducts with red light which are thermally stable at room temperature and that can be photochemically erased. Dependent on the light composition both properties can be realized in the same sample material. This feature may bridge the gap between information processing and short-term and long-term storage of information with BR.

Bacteriorhodopsins↗
PubMed1991

Indomethacin ion-selective electrode based on a bis(triphenylphosphoranylidene)ammonium-indomethacin complex.

The properties of an ion-selective electrode for indomethacin based on the bis(triphenylphosphoranylidene)ammonium-indomethacin ion-pair complex in a poly(vinyl chloride) membrane are described. The detection limit found for indomethacin at pH 7.0 was 2 x 10(-5) mol l-1 (6.4 micrograms ml-1) and the linear range was determined to be between 5 x 10(-5) and greater than or equal to 10(-3) mol l-1. The selectivity coefficients observed for organic anions of different lipophilicity were 10(-1.1) for naproxenate, 10(-0.75) for salicylate and less than 10(-4) for tartrate. Cationic organic and inorganic ions showed negligible interference with selectivity coefficients of less than 10(-4). A strong electrode response was observed for the inorganic anions IO4-, ClO4- and SCN- with selectivity coefficients of up to 10(+0.9). Very low detection limits, e.g., 6 x 10(-7) mol l-1 (60 ng ml-1) for ClO4- were found. The selectivity for inorganic anions was close to the Hofmeister series.

Electrodes↗
PubMed1991

Determination of ethaverine and papaverine using ion-selective electrodes.

Ion-selective poly(vinyl chloride) membrane electrodes for the opium alkaloids papaverine and ethaverine are presented. The electrode membranes contain ion pairs of the alkaloids with the anionic counter ion tetraphenylborate. The detection limits for all electrodes were approximately 2 x 10(-6) mol dm-3 at pH 5.0 in 100 mmol dm-3 buffered solutions and the measured slopes were close to the values theoretically expected. The selectivity coefficient observed for the ethaverine-tetraphenylborate electrode is 10(-1.1) with respect to papaverine. The suitability of the membranes for single-use electrodes is discussed.

Electrochemistry↗
PubMed1988

The cDNA clone for strictosidine synthase from Rauvolfia serpentina. DNA sequence determination and expression in Escherichia coli.

The cDNA clone for strictosidine synthase, the enzyme which catalyzes the stereospecific condensation of tryptamine with secologanin to form the key intermediate in indole alkaloid biosynthesis, strictosidine, has been identified with a synthetic oligodeoxynucleotide hybridization probe in a lambda gt11 cDNA library of cultured cells of Rauvolfia serpentina. The DNA has been sequenced, revealing an open reading frame of 1032 base pairs encoding 344 amino acids. The sequence of 60 nucleotides in the 5'-flanking region has been determined by primer extension analysis. The encoded protein has been expressed in E. coli DH5 as detected by immunoblotting of protein extracts with antibodies raised against the native enzyme.

Amino Acid Sequence↗
PubMedDate not supplied

Nanodissection and noncontact imaging of plasmid DNA with an atomic force microscope.

In this paper we report that a combination of noncontact and contact atomic force microscopy is a convenient and reliable method for imaging and dissecting single plasmid deoxyribonucleic acid (DNA) strands on mica at ambient conditions without leaving feedback and without damage to the scanning tips. The width and thickness measured at different points of the DNA strands agree with literature data and are the same before and after dissection.

DNA↗
PubMedDate not supplied

Improvement of DNA-visualization in dynamic mode atomic force microscopy in air.

Near-contact mode atomic force microscopy (AFM) imaging leads to sharper representations of DNA double strands on mica imaged at ambient conditions compared with noncontact mode AFM. Phase shift was used for feedback control yielding height information using a simple model calculation. No contact between tip and sample occurs. Measured DNA widths were up to four times smaller than measured with the same AFM tip in noncontact mode at ambient condition.

Animals↗