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K L Manchester

Publications and source records attributed to K L Manchester.

At least 19 recordsLinked to original sources
PubMed2001

Catalysis of guanine nucleotide exchange on eIF2 by eIF2B: can it be both a substituted enzyme and a sequential mechanism?

There are conflicting reports over the question of whether the displacement by GTP of GDP bound to eIF2 catalyzed by eIF2B follows a substituted enzyme mechanism, as is believed to be the case for other guanine nucleotide exchange factors, or is a sequential mechanism. Analysis of data recently provided by Williams et al. (J. Biol. Chem. 276, 24697-24703, 2001) showing displacement by eIF2B of GDP bound to eIF2 in the absence of displacing nucleotide appears to offer a way of resolving the dispute and suggests that both mechanisms may be operative.

Catalysis↗
PubMed2001

Antoine Béchamp: pere de la biologie. Oui ou non?

There is an alternative medicine lobby that, in conjunction with antivivisectionists, believes Louis Pasteur to have been a fraud [R. Bottomley's You Don't Have to Feel Unwell! (Newleaf, 1994) is a recent example]. They frame their accusations around a rivalry between Pasteur and a contemporary, Antoine Béchamp, from whom they suggest Pasteur stole his ideas and then distorted them for his own purposes. This article explores some aspects of the controversies between Béchamp and Pasteur.

Biology↗
PubMed2000

Biochemistry comes of age: a century of endeavour.

Biochemistry began as an identifiable, separate discipline at the beginning of the twentieth century. This article traces its early development, focusing on three pioneers--Eduard Buchner, Arthur Croft Hill and Arthur Harden.

Biochemistry↗
PubMed1999

The use of poly(2-acrylamido-2-methyl-1-propanesulfonic acid) polymers as spacers for isotachophoresis in sieving gel matrices.

The electric field strength gradients generated in isotachophoresis (ITP) may be used for the separation of biomolecules. Poly(2-acrylamido-2-methyl-1-propanesulfonic acid) (polyAMPS) polymers of a uniform distribution of molecular mass were synthesized and used as novel spacers in ITP. Since these polymeric spacers are strongly acidic species, their ionic charges remain constant over a wide pH range, so that their ionic mobilities are governed solely by their molecular masses and not by the pH of the milieu. A modification of ITP known as telescope electrophoresis was used to separate a number of acidic dyes of varying ionic mobility, using polyAMPS polymers as spacers. The resolution obtained was superior to that obtained by polyacrylamide gel electrophoresis (PAGE), due to the focusing effect of the electric field strength gradient. Since these novel polymeric spacers are designed to operate within sieving medium, it was decided to test their suitability for the separation of DNA molecules. DNA molecules up to 1000 bp long were successfully resolved, with a similar resolution to that obtained with conventional PAGE.

Coloring Agents↗
PubMed1998

Synthesis of zwitterionic acrylic acid buffers for isoelectric focusing in immobilized pH gradients.

A range of zwitterionic acrylic acid derivatives, buffering in the neutral and basic pH ranges, have been synthesized by the Mannich reaction of malonic acid, formaldehyde and a secondary amine. These compounds include 2-(4-morpholinomethyl)propenoic acid pK2 7.59 +/- 0.03 (23 degrees C), 2-[bis(2-hydroxyethyl)aminomethyl]propenoic acid pK2 approximately 8.6 (20 degrees C), 2-[bis(2-hydroxypropyl) aminomethyl]propenoic acid PK2 approximately 8.7 (20 degrees C), 2-[N-(2-hydroxyethyl)-N-methylaminomethyl]propenoic acid pK2 9.22+/-0.08 (22 degrees C), 2-[N-ethyl-N-(2hydroxyethyl)aminomethyl]-propenoic acid pK2 approximately 9.6 (20 degrees C), and 2-[4-(2-carboxyprop-2-enyl)piperazinylmethyl]propenoic acid, which has a sigmoidal buffering profile over the pH range 3-10. These zwitterionic acrylic acid buffers were successfully copolymerized with acrylamide to prepare immobilized pH gradients (IPGs) in the neutral to alkaline portion of the pH range. Bovine erythrocyte carbonic anhydrase isozymes were resolved on a pH 5-8 IPG prepared using 2-[4-(2-carboxyprop-2-enyl)piperazinylmethyl]propenoic acid as the immobilized buffer, and horse heart myoglobin was focused on pH 7.1-8.1 and pH 7.5-7.7 IPGs, using 2-(4-morpholinomethyl)propenoic acid as the immobilized buffer. In both cases the pK 9.3 Immobiline compound was used as the strongly basic titrant. These new compounds, besides possessing more hydrophilic residues than the corresponding commercial basic acrylamido buffers (Immobilines), resist hydrolysis at alkaline pH values.

Acrylates↗
PubMed1997

Catalysis of guanine nucleotide exchange on eIF-2 by eIF-2B: is it a sequential or substituted enzyme mechanism?

The mechanism of action of the eukaryotic initiation factor eIF-2B in catalyzing the exchange of guanine nucleotides bound to eIF-2 is uncertain--evidence having been adduced for a sequential mechanism and for a substituted enzyme mechanism. Data purporting to support a substituted enzyme mechanism have been analysed and shown to be ambiguous and equally consistent with a sequential mechanism. Suitable rate constants for a sequential mechanism involving the transient formation of the quaternary complex eIF-2.eIF-2B.GDP.GTP are suggested.

Catalysis↗
PubMed1997

Binding constants in the formation of mammalian protein synthesis initiation complexes and the role of mRNA.

The findings of Parkhurst et al. (Biochemistry 33, 15168-15177:1994) that a 10-mer oligoribonucleotide containing the AUG triplet enhances the binding of the eIF-2 x Met-tRNAi complex to the 40S ribosomal subunit are questioned on the basis of a re-evaluation of their calculations. It is not possible to conclude, as they did, that addition of the AUG-containing oligonucleotide produces an exceptionally large increase (as judged by the magnitude of the coupling free energy) in the binding of the eIF-2 x Met-tRNAi complex to the 40S subunit, or that their results are more consistent with internal initiation than with the scanning initiation model.

Animals↗
PubMed1996

Exploring the gene with X-rays.

X-rays were discovered 100 years ago. Their effects on living cells and on mutation were quickly recognized. This article discusses a classic paper that sought to use X-rays to estimate the size of the gene and question mechanics to explain its impressive stability.

Chemical Phenomena↗
PubMed1995

Membrane potential of rat hepatoma cells in culture: influence of factors affecting amino acid transport.

The effect has been studied of various media, hormones and of amino acids on the membrane potential of rat hepatoma cells in culture measured by microelectrode impalement. Cells in Eagle's minimal essential medium plus 5% serum had a value which varied daily from about 5-8 mV, inside negative. The membrane potential of rat hepatocytes was measured to be 8.7 +/- 0.2 mV, inside negative. The membrane potential of the hepatoma cells was decreased by insulin and increased by glucagon. Membrane potential was unaffected by change of medium to Hanks' or Earle's balanced salt solutions or deprivation of serum. It was, however, reduced in cells in phosphate-buffered saline and by reduction of pH. The former effect was shown to be due to the higher [Na+] of phosphate-buffered saline as opposed to the other media. Addition of alanine, glycine, serine, proline and methylaminoisobutyrate all reduced membrane potential by 2-3 mV. Smaller decreases were seen with methionine, leucine and phenylalanine, but none with glutamine, threonine, BCH (2-aminonorborane-2-carboxylic acid) and D-alanine. The results are compared with the effects of similar conditions on aminoisobutyrate uptake. Whilst there was a correlation under some conditions there was not under others. It is concluded that for the hepatoma cells factors additional to the membrane potential must exert some influence on the capacity for amino acid transport.

Amino Acids↗
PubMed1995

Louis Pasteur (1822-1895)--chance and the prepared mind.

Louis Pasteur is often regarded as one of the founders of microbiology, and is chiefly famous for his discovery of the role that microorganisms play in health and disease. However, he was trained as a physical scientist, and his research began with the discovery of the stereoisomerism of the different forms of tartaric acid. As a biotechnologist, he is remembered for his studies on the nature of fermentation, and for his rebuttal of the theory of spontaneous generation.

Biochemistry↗
PubMed1994

The 9-kDa calbindin gene of Rousettus aegyptiacus: its identification and isolation from a genomic library.

A genomic library of the fruit bat (Rousettus aegyptiacus) was constructed in lambda phage gt11. The titre of the library was determined to be 2 x 10(5) pfu/ml. The genomic library was amplified and the titre of the amplified library increased 300-fold to 7 x 10(7) pfu/ml. The library was screened by in situ hybridization techniques using a fragment of the mouse 9-kDa calbindin cDNA as a probe. Screening of 10(5) plaques yielded a positive clone. Three additional rounds of screening were performed to purify the positive. Lambda phage DNA was isolated from the positive clone and restriction digest analysis, followed by hybridization studies, was performed on these digests in order to determine the location of the bat 9-kDa calbindin gene in the insert of the lambda phage vector. Restriction maps so derived were interpreted from the published sequence for the rat 9-kDa calbindin gene and indicate the successful isolation of the 9-kDa calbindin gene of Rousettus aegyptiacus.

Animals↗