Search PubMed⌕ Search

Biomedical subjects

M K Johnson

Publications and source records attributed to M K Johnson.

At least 199 records · Page 11Linked to original sources
PubMed1985

The iron-sulfur cluster composition of Escherichia coli nitrate reductase.

Nitrate reductase from Escherichia coli has been investigated by low-temperature magnetic circular dichroism and electron paramagnetic resonance (EPR) spectroscopies, as well as by Fe-S core extrusion, to determine the Fe-S cluster composition. The results indicate approximately one 3Fe and three or four [4Fe-4S]2+,1+ centers/molecule of isolated enzyme. The magnetic circular dichroism spectra and magnetization characteristics show the oxidized and reduced 3Fe and [4Fe-4S] centers to be electronically analogous to those in bacterial ferredoxins. The form and spin quantitation of the EPR spectra from [4Fe-4S]1+ centers in the reduced enzyme were found to vary with the conditions of reduction. For the fully reduced enzyme, the EPR spectrum accounted for between 2.9 and 3.5 spins/molecule, and comparison with partially reduced spectra indicates weak intercluster magnetic interactions between reduced paramagnetic centers. In common with other Fe-S proteins, the 3Fe center was not extruded intact under standard conditions. The results suggest that nitrate reductase is the first example of a metalloenzyme where enzymatic activity is associated with a form that contains an oxidized 3Fe center. However, experiments to determine whether or not the 3Fe center is present in vivo were inconclusive.

Circular Dichroism↗
PubMed1985

Quarter field resonance and integer-spin/half-spin interaction in the EPR of Thermus thermophilus ferredoxin. Possible new fingerprints for three iron clusters.

We describe two new characteristics of the EPR of the seven-iron containing ferredoxin from Thermus thermophilus. First, the reduced state of the 3Fe center, which has traditionally been considered to be EPR-silent, has been found to exhibit a delta m = 4 transition, which is unique for Fe-S centers. This signal is similar to that of high-spin Fe2+-EDTA and supports the suggestion that the ground electronic state of the 3Fe cluster is S = 2. Second, we have recorded the EPR spectrum of the fully reduced protein at 9 and 15 GHz and found that changes occur in the signal which are consistent with a weak electronic spin-spin interaction between the [4Fe-4S]+ (S = 1/2) and the reduced 3Fe center. A theoretical explanation is given for the observation of interaction signals with constant effective g values.

Electron Spin Resonance Spectroscopy↗
PubMed1985

Low temperature magnetic circular dichroism spectroscopy as a probe for the optical transitions of paramagnetic nickel in hydrogenase.

A partially-purified sample of hydrogenase from Methanobacterium thermoautotrophicum (delta H strain) has been investigated by optical absorption, magnetic circular dichroism and electron paramagnetic resonance spectroscopy. Variable temperature magnetic circular dichroism studies reveal, for the first time, the optical transitions associated with the Ni(III) center in the oxidized enzyme. Low temperature magnetic circular dichroism spectroscopy provides a new method of assessing both the coordination environment of Ni in hydrogenase and the appropriateness of inorganic model complexes.

Circular Dichroism↗
PubMed1985

Do alcoholic Korsakoff's syndrome patients acquire affective reactions?

In this study we report two experiments that investigate the acquisition of affective reactions. In Experiment 1, unfamiliar melodies were played to Korsakoff's syndrome patients and alcoholic and nonalcoholic control subjects who were matched with them according to age and education. Following a retention interval of 5 min, subjects received a preference test on old and new melodies. Korsakoff's syndrome patients showed the same increase in preference for old melodies as a consequence of prior exposures as control subjects did, but their recognition of melodies was significantly impaired in comparison with controls. In Experiment 2, the same subjects saw photographs of two men. Fictional biographical information depicted one as a "good guy" and the other as a "bad guy." After a retention interval of approximately 20 days, Korsakoffs recalled virtually none of the biographical information; however, 78% preferred the good guy, and impression ratings were less favorable for the bad guy. Korsakoff patients developed preferences and impressions even though they did not have voluntary access to the information on which the preferences were based. However, their impression ratings were less extreme than those of controls. The pattern of results of the two studies is discussed in terms of Johnson's (1983) MEM model of memory.

Adult↗
PubMed1985

Confusions between memories for performed and imagined actions: a developmental comparison.

Children are often assumed to be more confused than adults are about the origin of self-generated memories (e.g., what they did or thought). The present experiments showed evidence in support of this assumption but only under some circumstances. In Experiment 1, 6- and 9-year-olds were as good as adults in distinguishing what they did from what they saw someone else do. However, children had particular trouble distinguishing what they did from what they imagined doing. Confusion between performed and imagined actions was evident across a range of actions. Clustering data also showed that information about origin is part of the memory for an event; all subjects recalled actions according to who performed what action (Experiment 1). Further, the presence of person categories as a basis for organization reduced clustering based on action class more for children than for adults (Experiment 1 vs. 2). Collectively, these findings indicate that children become sensitive to some distinctions in memories sooner than they do to others.

Child↗
PubMed1984

Characterization of the Fe-S cluster in aconitase using low temperature magnetic circular dichroism spectroscopy.

Beef heart aconitase has been studied by low temperature magnetic circular dichroism (MCD) spectroscopy in the wavelength region 300 to 1900 nm. Together with parallel electron paramagnetic resonance and activity measurements, these data enable correlations between Fe-S cluster-type and enzymic activity in aconitase. In samples not exposed to extraneous Fe, the Fe-S cluster in aconitase exhibits the characteristic properties of a 3Fe center in both the as isolated and dithionite-reduced states. On the basis of the detailed form of the low temperature MCD spectra, three types of 3Fe center can be distinguished in biological samples. These are typified by the 3Fe centers in aconitase, Desulfovibrio gigas FdII, and Azotobacter crooccocum Fd. In aconitase, maximal enzymic activity is found to be associated with the build-up of [4Fe-4S]2+ clusters in good agreement with the Mössbauer studies of Kent et al. (Kent, T. A., Dreyer, J. L., Kennedy, M. C., Huynh, B. H., Emptage, M. H., Beinert, H., and Münck, E. (1982) Proc. Natl. Acad. Sci. U. S. A. 79, 1096-1100). However, significant catalytic activity (approximately 60%) was obtained by reduction of the 3Fe center with dithionite in the absence of added Fe. The form and intensity of the resultant MCD spectrum are consistent with the majority of the Fe being in the form of reduced 3Fe clusters. The possibility that a reduced 3Fe cluster is capable of promoting partial catalytic activity in aconitase is discussed in light of these results.

Aconitate Hydratase↗
PubMed1984

Adherence of Staphylococcus aureus to rabbit corneal epithelial cells.

A system for the in vitro study of adherence of Staphylococcus aureus to rabbit corneal epithelial cells has been developed. Bacteria isolated from normal eyes showed the same adherence capacity, on the average, as bacteria from infected eyes. Adherence was not inhibited by preincubation of bacteria with bovine serum albumin, D-mannose, or N-acetyglucosamine.

Adhesiveness↗
PubMed1984

Dreams and reality monitoring.

Experiment 1 tested the counterintuitive prediction that memories for one's own dreams should not be particularly easy to discriminate from memories for someone else's dreams. Pairs of people reported dreams to each other that they had either dreamed, read, or made up the night before. On a test requiring subjects to discriminate events they had reported from those reported by their partner, subjects had more difficulty with real dreams than with dreams they read or made up. Experiment 2 provided evidence that real dreams do not simply produce overall weaker memories; the deficit for dreams was eliminated with more time to respond and with more detailed cues. In addition, subjects' ratings of various characteristics of their memories (e.g., vividness, personal relevance) indicated that dreams were not generally weaker or impoverished. The results are interpreted within the framework for reality monitoring described by Johnson and Raye (1981): Memories for real dreams are proposed to be deficient in conscious cognitive operations that help identify the origin of information generated in a waking state. At the same time, real dreams are embedded in a network of supporting memories that can be drawn on for reality monitoring decisions under appropriate circumstances. Finally, a comparison of recognition and recall indicated that dreams may leave persisting memories that are difficult to access via free recall.

Adult↗
PubMed1983

Resonance Raman studies of beef heart aconitase and a bacterial hydrogenase.

The resonance Raman (RR) spectra of beef heart aconitase and of an air-stable hydrogenase from Desulfuvibrio desulfuricans, as isolated, are characteristic of 3Fe centers. Activation of aconitase by Fe(II) addition converts the RR spectrum to one characteristic of [4Fe-4S]2+ clusters. Analytical data on aconitase, as isolated, confirms the recent finding (Beinert, H., Emptage, M. H., Dreyer, J.-L., Scott, R. A., Hahn, J. E., Hodgson, K. O., and Thomson, A. J. (1983) Proc. Natl. Acad. Sci. U. S. A. 80, 393-396) of a [3Fe-4S] stoichiometry. The RR spectra of 3Fe centers from aconitase, and the hydrogenase, as well as from several bacterial ferredoxins, conform to the pattern expected for a cube-derived [3Fe-4S] cluster. Perceptible differences are observed among the spectra, which can be explained in terms of differences among the terminal ligands, perhaps limited to their conformations. In the case of aconitase and hydrogenase, frequency shifts suggest additional alterations in the terminal Fe-S bond angles and/or slight differences in core geometry.

Aconitate Hydratase↗
PubMed1983

Age-related changes in confusion between memories for thoughts and memories for speech.

The present experiments compared people's abilities to make decisions about the origin of their memories. Experiment 1 demonstrated that 6-year-olds were as good as 17-year-olds in discriminating memories originating from what they said earlier (self-generations) from memories of what another person said earlier (external presentations). However, in both experiments 1 and 2, 6-year-olds were not as good at discriminating what they had said earlier from what they had only thought. The possibility that younger children simply have more difficulty distinguishing between memories originating from the same class, internal or external, was ruled out because 6-year-olds performed as well as 9-year-olds when differentiating between memories from 2 external sources (experiment 2). Nor could their difficulty be attributed to a general problem in distinguishing memories for their thoughts from any other class of memories because they were at no disadvantage in discriminating their earlier thoughts (words they imagined themselves saying) from words someone else said (experiment 2). Our findings suggest that some distinctions, self versus other, emerge as cues in memory sooner than other distinctions, thoughts versus actions.

Adolescent↗
PubMed1982

The hydrophobic character of thiol-activated cytolysins.

Hydrophobic chromatography on phenyl-Sepharose has revealed the decidedly hydrophobic character of several members of the group of cytolytic proteins termed ;thiol-activated'. Pneumolysin, alveolysin, cereolysin, and streptolysin O were found to be equally hydrophobic, as were the oxidized and reduced forms of alveolysin. Hydrophobic chromatography has been utilized in the development of an improved procedure for the purification of pneumolysin.

Bacterial Proteins↗
PubMed1982

Resonance Raman and electron paramagnetic resonance studies on oxidized and ferricyanide-treated Clostridium pasteurianum ferredoxin. Vibrational assignments from 34S shifts and evidence for conversion of 4 to 3 iron-sulfur clusters via oxidative damage. Vibrational assignments from 34S shifts and evidence for conversion of 4 to 3 iron-sulfur clusters via oxidative damage.

Resonance Raman spectra are reported for oxidized ferredoxin from Clostridium pasteurianum and for protein reconstituted with 34S2-, using 4579 A laser excitation. The spectra are of much higher quality than that previously reported, and the 34S shifts provide assignments of the Fe-S modes. After treatment with ferricyanide, the resonance Raman spectrum closely resembles that of the [3Fe-3S] protein, ferredoxin II from D. gigas; the 34S shifts aid in assignments of the [3Fe-3S] modes. The epr signal associated with the [3Fe-3S] cluster (g = 2.01) corresponds maximally to 0.80 spin/molecule. Anaerobic addition of excess sulfide to the reduced, ferricyanide-treated protein regenerates a [4Fe-4S]1+ epr spectrum, equivalent in intensity to the [3Fe-3S] signal. The ubiquitous occurrence of a g = 2.01 signal in preparations of [4Fe-4S] proteins can be attributed to variable amounts of [3Fe-3S] cluster generated by adventitious oxidation. The ready conversion of [4Fe-4S] to [3Fe-3S] clusters in C. pasterianum raises the possibility that some [3Fe-3S] proteins may actually arise by oxidative damage of [4Fe-4S] proteins during aerobic isolation.

Clostridium↗