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K Moreton

Publications and source records attributed to K Moreton.

18 recordsLinked to original sources
PubMed2001

The A245K mutation exposes another stage of the bacterial L-lactate dehydrogenase reaction mechanism.

The A245K mutant of Bacillus stearothermophilus L-lactate dehydrogenase has been expressed in Escherichia coli and purified. A qualitative change in the reaction mechanism prior to the hydride transfer step in the reverse direction in the mutant is revealed. Both transient and steady state characteristics of the mutant are presented and show in contrast to the wild-type enzyme where a rearrangement of an enzyme-NADH-pyruvate complex is rate-limiting that in the mutant the rearrangement is much faster and hydride transfer is the first slow step. The steady state is limited by a new second slower conformation change involving an NAD+ complex. The mutation may provide a valuable framework for inhibitor and drug design research.

Alanine↗
PubMed1995

Protein expression of the alpha, gamma, delta and epsilon subspecies of protein kinase C changes as C6 glioma cells become contact inhibited and quiescent in the presence of serum.

Total protein kinase C (PKC) activity and protein expression of the alpha and delta subspecies of PKC increases markedly as C6 glioma cells grow from low cell density to the contact-inhibited quiescent state (also known as G(o)) in the presence of serum. At the same time protein expression of PKC subspecies gamma and epsilon decreases while the beta I, beta II, iota and zeta subspecies did not change. Serum deprivation of growing C6 glioma cells does not induce the same changes in PKC subspecies protein expression. The findings support the growing view that there are significant differences between the G(o) states brought about by contact inhibition or serum deprivation.

Blood↗
PubMed1992

Polymeric ovotransferrin and its ability to bind and deliver iron to chick-embryo red blood cells.

Polymeric forms of hen ovotransferrin have been obtained by storage at 4 degrees C for 5 years or at 57 degrees C for 14 days and fractionated as dimers and tetramers by gel filtration on Sephadex. The ability of tyrosine to undergo nitration was reduced in the tetrameric protein, so that one could hypothesize that dityrosine formation is somewhat responsible of such polymerization process. Experimental data on the biological functionality showed that: i) dimeric ovotransferrin was able neither to bind nor to deliver iron; ii) tetrameric ovotransferrin was able to bind but not to deliver iron.

Animals↗
PubMed1990

Binding and iron delivering of monoferric ovotransferrins to chick-embryo red blood cells (CERBC).

1. Both monoferric forms of OTf, each of about 80 kDa, bound to CERBC enough tightly, but at a lesser extent with respect to Fe2OTf with a Bmax in the order: 59Fe2OTf greater than OTf59FeC much greater than 59FeNOTf. 2. Fe2OTf competed, in equimolar ratio, with 59FeNOTf or OTf59FeC, lowering the Bmax value; a 10-fold molar excess of Fe2OTf almost abolished the binding of both labelled monoferric forms. Apo-OTf did not compete with the monoferric forms for binding to CERBC. Iron-saturated N- or C-terminal OTf half-molecules, each of about 40 kDa, were unable to displace the monoferric form. By contrast, the mixture of both half-molecules gave results very similar to Fe2OTf. 59FeNOTf and OTf59FeC were displaced by a molar excess of both unlabelled monoferric forms. 3. Uptake experiments showed that both monoferric forms of OTf were less effective in delivering iron to CERBC with respect to the diferric form, but, nevertheless, there was still an appreciable iron uptake which paralleled the binding behaviour, being the C-form slightly more efficient than the N-form.

Animals↗
PubMed1989

Preliminary crystallographic studies on duck ovotransferrin.

Crystals of duck ovotransferrin and duck apo-ovotransferrin have been grown from polyethylene glycol solutions. For both crystals, the space group is P2(1)2(1)2(1), the unit cell dimensions for the ovotransferrin are a = 49.6 A, b = 85.6 A, c = 178.7 A and for the apo-ovotransferrin a = 77.6 A, b = 98.8 A, c = 127.0 A, giving four molecules in the unit cell.

Animals↗
PubMed1989

Binding of various ovotransferrin fragments to chick-embryo red cells.

1. The ability of N- and C-terminal half-molecule fragments of hen ovotransferrin to interact with chick red blood cells (CERBC) has been studied under conditions that allow binding of the transferrin to transferrin receptors to take place, but not the delivery of iron to the cell. Two kinds of half-molecule fragments were used: (a) those which can associate with one another to give a dimer resembling native transferrin and (b) those which cannot associate in this way because they lack a few amino acid residues from their C-terminal ends. 2. Neither N nor C half-molecules alone can bind to the CERBC, but, when both are present, tight binding occurs. 3. Whether or not the half-molecules can associate with one another makes little difference to receptor binding. 4. Given that one of the half-molecules is iron-saturated, the presence or absence of iron in the contralateral half-molecule again makes little difference to receptor binding.

Animals↗
PubMed1988

The dimerization of half-molecule fragments of transferrin.

Partial proteolysis was used to prepare half-molecule fragments of hen ovotransferrin. N-Terminal and C-terminal fragments associate to form an N-terminal fragment-C-terminal fragment dimer. Variant forms of the N- and C-terminal fragments can be prepared in which a few amino acid residues are lacking from the C-terminal ends of the fragments. These variant fragments are partially or completely unable to associate; the suggestion that the molecular recognition sites are located in these C-terminal stretches of the N-terminal half-molecule (320-332) and of the C-terminal half-molecule (683-686) is in agreement with X-ray-crystallography data for human lactotransferrin [Anderson, Baker, Dodson, Norris, Rumball, Waters & Baker (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 1769-1773].

Amino Acid Sequence↗
PubMed1985

Selective reduction of a disulphide bridge in hen ovotransferrin.

Brief treatment of iron-saturated hen ovotransferrin with dithiothreitol selectively cleaves the disulphide bridge between residues 478 and 671, which is in the C-terminal domain of the protein. The reduced alkylated protein is less stable than the native protein, and its iron-binding properties are different. A fluorescent derivative was prepared by coupling N-iodoacetyl-N'-(5-sulpho-1-naphthyl)ethylenediamine to the thiol groups.

Alkylation↗
PubMed1984

Value of serum diazepam and nordiazepam measurements in anxious patients.

The relationship between the dose and anxiolytic effects of diazepam and the serum concentrations of diazepam and nordiazepam were examined in groups of acutely and chronically anxious patients. The results showed a significant correlation between dose and serum nordiazepam concentrations after short-term (14-day) administration, but no significant association between clinical symptoms of anxiety and serum diazepam and nordiazepam. We conclude that the main value of serum benzodiazepine measurements in anxious patients in assessing compliance, particularly in patients suspected of taking more than the recommended dose. Serum nordiazepam is a more consistent index of dosage after chronic therapy than serum diazepam because it has a longer elimination half-time.

Adult↗
PubMed1982

The effect of salt concentration on the iron-binding properties of human transferrin.

The salt dependence of the iron-binding properties of transferrin was studied by urea/polyacrylamide-gel electrophoresis. The distribution of iron between the N-terminal and C-terminal binding sites under equilibrium conditions and the rates of release of iron from the two sites were studied. It was found that salt increases the thermodynamic stability of iron binding in the N-terminal site relative to the C-terminal site. Similar behaviour is observed for the kinetics of iron release, where salt retards the rate of removal of iron from the N-terminal site but facilitates removal from the C-terminal site.

Binding Sites↗
PubMed1982

A variant of human transferrin with abnormal properties.

Screening of human serum samples by polyacrylamide-gel electrophoresis in the presence of 6 M-urea revealed an individual who is heterozygous for a variant transferrin. The variant transferrin is able to bind two atoms of iron, but the iron in the C-terminal binding site is bound abnormally, as judged by its spectral properties, and is dissociated from the protein on electrophoresis in the presence of 6 M-urea. The iron-free C-terminal domain of the variant protein is less stable than normal to thermal and urea denaturation. Structural changes in the variant protein have not yet been characterized.

Amino Acids↗
PubMed1980

The distribution of iron between the metal-binding sites of transferrin human serum.

The Makey & Seal [(1976) Biochim. Biophys. Acta 453, 250--256] method of polyacrylamide-gel electrophoresis in buffer containing 6 M-urea was used to determine the distribution of iron between the N-terminal and C-terminal iron-binding sites of transferrin in human serum. In fresh serum the two sites are unequally occupied; there is preferential occupation of the N-terminal site. On incubation of the serum at 37 degrees C the preference of iron for the N-terminal site becomes more marked. On storage of serum at -15 degrees C the iron distribution changes so that there is a marked preference for the C-terminal site. Dialysis of serum against buffer at pH 7.4 also causes iron to be bound much more strongly by the C-terminal than by the N-terminal site. The original preference for the N-terminal site can be resroted to the dialysed serum by addition of the diffusible fraction.

Binding Sites↗
PubMed1978

The iron-binding properties of hen ovotransferrin.

1. The distribution of iron between the two iron-binding sites in partially saturated ovotransferrin was studied by labelling with 55Fe and 59Fe and by gel electrophoresis in a urea-containing buffer. 2. When iron is added in the form of chelate complexes at alkaline pH, binding occurs preferentially at the N-terminal binding site. In acid, binding occurs preferentially at the C-terminal site. 3. When simple iron donors (ferric and ferrous salts) are used the metal is distributed at random between the binding sites, as judged by the gel-electrophoresis method. The double-isotope method shows a preference of ferrous salts for the N-terminal site. 4. Quantitative treatment of the results of double-isotope labelling suggests that in the binding of iron to ovotransferrin at alkaline pH co-operative interactions between the sites occur. These interactions are apparently absent in the displacement of copper and in the binding of iron at acid pH.

Binding Sites↗
PubMed1972

Narcosis therapy.

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Depression↗