Search results for "rase"

Comparison of the interaction of cobalt bovine carbonic anhydrase II with acetazolamide and methazolamide and the reaction of apoenzyme with cobalt(I…

2003

The metalloenzyme carbonic anhydrase (CA) is an attractive choice for a research-based bioinorganic laboratory course. In this project the interaction of cobalt bovine carbonic anhydrase II (CoBCAII) with acetazolamide and methazolamide and the reaction of apoenzyme with cobalt(II) complexes of acetazolamide and methazolamide is studied by UV-visible spectroscopy. Prior to this spectroscopic study students are given native BCAII, and they prepare apoBCAII and CoBCAII. A major aim is to provide experience in handling metalloproteins and in the study of metal complexes-protein interactions.

The Mechanism of Action of Carbonic Anhydrase

1990

A survey of the structure-function relationship in the enzyme carbonic anhydrase is presented. The coordination number around the metal ion in derivatives inhibited with some anions is stressed as well as the role of the cavity in determining the coordination number. Inferences on the enzyme-substrate interactions are presented.

Mannosyl transferases inSaccharomyces cerevisiae: Evidence for the occurrence of ectomannosyltransferase activity

1981

The subcellular distribution of mannosyltransferases inSaccharomyces cerevisiae was studied following the separation of the plasma membrane from other intracellular membranous systems. Most of the activity was linked to internal membranes, and the rest was located at the level of the plasma membrane. Yeast plasma membranes coated on their external face with concanavalin A when incubated with GDP-[U-14C]mannose incorporated 20% less [U-14C]mannose in glycoproteins and 110% more in glycolipids than plasma membranes alone. This suggested that part of the total mannosyltransferase activity of the plasma membrane is located on its outer surface. A significant incorporation of radioactive mannose…

Immediate early response of the marine sponge Suberites domuncula to heat stress: Reduction of trehalose and glutathione concentrations and glutathio…

1997

The marine sponge Suberites domuncula was used to identify early markers for thermal stress. Cubes from sponges have been kept for 30 min at 31°C (10 °C higher than the ambient temperature). After this treatment the sponge cubes were kept again at 21°C. To demonstrate that the animals reacted to the elevated temperature, the expression of heat shock protein (HSP) was determined. Using an antibody raised against HSP70, it was found by Western blotting that the animals specifically express a 45 kDa polypeptide after heat treatment. It was shown that even after 10 min of heat treatment the steady-state concentration of trehalose drops by 40% from a base level of 13 nmol/mg protein. The activit…

Chemie an starren Grenzflächen, 10. Kinetisches Verhalten einiger an „Isothiocyanato-Aerosil” immobilisierter Esterasen – Analogversuche mit Insulin

1988

Amino-Aerosil wird mit Thiophosgen nach Gl. (1) in Isothiocyanato-Aerosil 2 (NCS-Aerosil) ubergefuhrt. Trypsin, Cholinesterase und Acetylcholinesterase werden an 2 nach Gl. (2) immobilisiert. Die Aktivitat der immobilisierten Esterasen wird untersucht auf (a) Lagerstabilitat, (b) Temperaturbestandigkeit und (c) pH-Abhangigkeit. Das kinetische Verhalten der immobilisierten Enzyme wird verglichen mit dem der freien, aber mit n-Butylisothiocyanat behandelten Esterasen. Am Beispiel der Cholinesterase, die uber unterschiedliche Ankergruppen mit Aerosil verknupft wurde, wird der Einflus unterschiedlicher Spacergruppen auf die Hydrolysegeschwindigkeit studiert. Aerosile, die mit Acetylcholin-Analo…

Acetylcholine esterase: the structure.

1991

Biochemical Characterization of an Acetylcholine-hydrolyzing Enzyme from Bean Seedlings.

1980

An acetylcholine hydrolyzing enzyme was prepared and purified (40 times) from dwarf bean hypocotyl hooks. The purity of the enzyme was proved by polyacrylamide gel electrophoresis. The molecular weight of the enzyme was determined to be 65,000 daltons. Enzyme activity was the highest at pH 8.0 and between 30 and 36 C. The enzyme had an apparent affinity constant (K(m)) for acetylcholine of 460/micromolar. The affinity for substrate analogs increased from butyrylthiocholine to propionylthiocholine to acetylthiocholine. The enzyme activity was inhibited by choline, neostigmine, physostigmine, manganese, and calcium. Magnesium had no influence on the enzyme activity. We conclude that the enzym…

Cell wall-degrading enzymes produced in vitro by isolates of Phaeosphaeria nodorum differing in aggressiveness

2000

The relationships between in vitro production of cell wall-degrading enzymes and aggressiveness of three Phaeosphaeria nodorum isolates were investigated. When grown in liquid medium containing 1% cell wall from wheat leaves as the carbon source, the isolates secreted xylanase, α-arabinosidase, β-xylosidase, polygalacturonase, β-galactosidase, cellulase, β-1,3-glucanase, β-glucosidase, acetyl esterase and butyrate esterase. Time-course experiments showed different levels of enzyme production and different kinetics between isolates. A highly aggressive isolate produced more xylanase, cellulase, polygalacturonase and butyrate esterase than did the two weakly aggressive isolates. Xylanase was …

Self-instructed condensation of amino acids and the origin of biological information

1984

In contemporary cells biological information is largely stored in nucleic acids. Therefore, a prerequisite in many theories on the origin of cellular life is the pre-existence of self-replicating polynucleotides that had to be formed by abiotic processes on the prebiotic Earth. It is usually assumed that the spontaneous synthesis of a self-replicating polynucleotide could take place readily. However, serious stereochemical obstacles exist which make such a synthesis extremely improbable. Amino acids, on the other hand, which are abundantly formed in prebiotic simulation experiments, are relatively easily polymerized to macromolecules (protoproteins) that share with modern proteins many prop…

The gene encoding polyneuridine aldehyde esterase of monoterpenoid indole alkaloid biosynthesis in plants is an ortholog of theα/β hydrolase super fa…

2000

The biosynthesis of the anti-arrhythmic alkaloid ajmaline is catalysed by more than 10 specific enzymes. In this multistep process polyneuridine aldehyde esterase (PNAE) catalyses a central reaction by transforming polyneuridine aldehyde into epi-vellosimine, which is the immediate precursor for the synthesis of the ajmalane skeleton. PNAE was purified from cell suspension cultures of Rauvolfia serpentina. The N-terminal sequence and endoproteinase LysC fragments of the purified protein were used for primer design and for the amplification of specific PCR products leading to the isolation of PNAE-encoding cDNA from a R. serpentina library. The PNAE cDNA was fused with a C-terminal His-tag, …