scholarly journals pH-induced bistable dynamic behaviour in the reaction catalysed by glucose-6-phosphate dehydrogenase and conformational hysteresis of the enzyme

1989 ◽  
Vol 262 (3) ◽  
pp. 795-800 ◽  
Author(s):  
M A Aon ◽  
S Cortassa ◽  
J F Hervagault ◽  
D Thomas
Keyword(s):  
Dynamic Behaviour ◽  
Forward Direction ◽  
Memory Device ◽  
Reverse Direction ◽  
Protein Fluorescence ◽  
Ph Change ◽  
Before And After ◽  
Continuously Stirred Tank Reactor ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Intrinsic Protein Fluorescence

1. Bistable (multiple stationary states) dynamic behaviour in the activity of glucose-6-phosphate dehydrogenase that was subjected to successive pH change was demonstrated in an open continuously stirred tank reactor. Although the enzyme under study did not exhibit an autocatalytic effect and was homogeneously distributed, bistability was shown to occur. 2. The successive pH changes of the enzyme solution corresponded to a pH transition (8.3 in equilibrium 2), i.e. an acidification (forward direction) and an alkalinization (reverse direction). By use of intrinsic protein fluorescence methods, a glucose-6-phosphate dehydrogenase conformational hysteresis was shown to exist concomitant with the pH transition before and after enzyme injection into the reactor. 3. The results obtained suggest that the enzyme behaves, conformationally, as a memory device that stores information about its pH history (i.e. the enzyme records information in its structure about the environment to which it was previously exposed) and transduces it in a non-linear dynamic fashion, producing the bistable behaviour observed in the open reactor.

Thermotropic lipid and protein transitions in Chinese hamster lung cell membranes: relationship to hyperthermic cell killing

1983 ◽  
Vol 61 (6) ◽  
pp. 421-427 ◽  
Author(s):  
James R. Lepock ◽  
Kwan-Hon Cheng ◽  
Hisham Al-Qysi ◽  
Jack Kruuv
Keyword(s):  
Mammalian Cells ◽  
Plasma Membranes ◽  
Chinese Hamster ◽  
Cell Killing ◽  
Water Soluble ◽  
Growth Data ◽  
Protein Fluorescence ◽  
Polar Groups ◽  
Spin Resonance ◽  
Intrinsic Protein Fluorescence

Exposure of mammalian cells to hyperthermic temperatures (ca. 41–45 °C) appears to act as a direct or triggering effect to produce some later response such as cell death, thermotolerance, or heat-shock protein synthesis. The high activation energy of cell killing indicates that the direct effect of hyperthermia might be a thermotropic transition in some cellular component, for this particular response. Both hyperthermic survival and growth data imply that the temperature for the onset of hyperthermic cell killing is 40–41.5 °C for Chinese hamster lung V79 cells. Studies using the electron spin resonance label 2,2-dimethyl-5-dodecyl-5-methyloxazolidine-N-oxide and the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene show the existence of lipid transitions at approximately 7–8 and 23–36 °C (or a broad transition between these temperatures) in mitochondria and whole cell homogenates, that correlate well with changes in growth and hypothermic killing. No lipid transition was detected near 40–41.5 °C that could correlate with hyperthermic killing in either mitochondrial or plasma membranes, but measurements of intrinsic protein fluorescence and protein fluorophore to trans-paranaric acid energy transfer demonstrate the existence of an irreversible transition in protein structure or arrangement above ca. 40 °C in both mitochondrial and plasma membranes. This transition is due to protein rearrangement and (or) unfolding such that there is increased exposure of protein tryptophan and tyrosine residues to polar groups and to paranaric acid. The strength of the transition implies that a significant fraction of total membrane protein is involved in this transition, which may be analogous to the heat-induced denaturation of water-soluble proteins. This alteration in membrane structure above ca. 40 °C could cause many of the observed changes in plasma membrane and mitochondrial function, which may further be involved in cellular responses to hyperthermia.

scholarly journals Golgi-Colonnier method: correlation of the degree of chromium reduction and pH change with quality of staining.

1994 ◽  
Vol 42 (3) ◽  
pp. 393-403 ◽  
Author(s):  
A Angulo ◽  
J A Merchán ◽  
M Molina
Keyword(s):  
Chromium Reduction ◽  
Ph Change ◽  
Total Chromium ◽  
Chromium Species ◽  
Cell Staining ◽  
Before And After ◽  
Neuronal Proteins ◽  
Ph Increase

We examined the role of chromium reduction in the Golgi-Colonnier method, correlating the quality of neuronal impregnation with the levels of hexavalent (CrVI) and trivalent (CrIII) chromium in the tissue and in the chromation fluid (CF). The concentrations of both chromium species were assessed by measuring spectrophotometrically the CrVI before and after oxidizing the sample and by calculating the ratio of CrVI to total chromium (chromium ratio, CrR). The CrR was almost identical in the tissue and the CF, decreasing exponentially during chromation due to a progressive consumption of CrVI to form CrIII. Satisfactory cell impregnation was obtained only when the CrR was 0.45-0.7, regardless of other factors. The CrR values could be accurately predicted by the pH increase of the CF; this increase has proven to be a most reliable criterion to decide the endpoint of the chromation process. The dependence of cell staining on the [CrIII], together with the well-known ability of this species to bridge proteins, suggests that the key event for cell impregnation is the cross-linking of neuronal proteins by CrIII polymers.

Slow postcapillary pH changes in blood in anesthetized animals

1978 ◽  
Vol 45 (5) ◽  
pp. 674-680 ◽  
Author(s):  
A. Bidani ◽  
E. D. Crandall
Keyword(s):  
Blood Cells ◽  
Ph Change ◽  
Arterial Blood ◽  
Blood Ph ◽  
Pulmonary Capillary ◽  
Before And After ◽  
Equilibration Process ◽  
Contact Surfaces ◽  
Electrode Chamber

To investigate the hypothesis that blood pH and PCO2 continue to change after the blood leaves an exchange capillary, we used a rapidly responding, pressure-insensitive, stopped-flow pH electrode apparatus. Arterial blood from an anesthetized dog or cat is drawn through the apparatus into a syringe. Syringe movement is then suddenly stopped. Temperature and pH of the blood in the electrode assembly are continuously monitored, both before and after blood withdrawal ceases. Hemolysis was reduced by coating all blood contact surfaces with silicone and fasting the animal overnight, anesthetizing it with crystalline pentobarbital sodium, and allowing it to ventilate spontaneously. After stopping withdrawal, pH of blood in the electrode chamber continued to change, rising 0.01 unit with t1/2 of 4.4 s. After lysed blood was returned to the animal to provide carbonic anhydrase to the plasma, no pH change was seen after stopping the flow. The small pH rise occurring in arterial blood in vivo is probably due in large part to disequilibrium of [H+] between red blood cells and plasma at the end of the pulmonary capillary, the equilibration process being rate-limited by the extracellular CO2 hydration-dehydration reaction.

Sodium and potassium binding to parvalbumins measured by means of intrinsic protein fluorescence

1983 ◽  
Vol 749 (2) ◽  
pp. 185-191 ◽  
Author(s):  
Eugene A. Permyakov ◽  
Lina P. Kalinichenko ◽  
Vyacheslav N. Medvedkin ◽  
Edward A. Burstein ◽  
Charles Gerday
Keyword(s):  
Protein Fluorescence ◽  
Intrinsic Protein ◽  
Potassium Binding ◽  
Intrinsic Protein Fluorescence ◽  
Sodium And Potassium

scholarly journals Brain-wide mapping of water flow perception in zebrafish

2020 ◽  
Author(s):  
Gilles Vanwalleghem ◽  
Kevin Schuster ◽  
Michael A. Taylor ◽  
Itia A. Favre-Bulle ◽  
Ethan K. Scott
Keyword(s):  
Water Flow ◽  
Lateral Line ◽  
Reverse Flow ◽  
Forward Direction ◽  
Brain Regions ◽  
Reverse Direction ◽  
Departure Point ◽  
Forward Flow ◽  
Larval Zebrafish ◽  
The Brain

AbstractInformation about water flow, detected by lateral line organs, is critical to the behavior and survival of fish and amphibians. While certain specific aspects of water flow processing have been revealed through electrophysiology, we lack a comprehensive description of the neurons that respond to water flow and the network that they form. Here, we use brain-wide calcium imaging in combination with microfluidic stimulation to map out, at cellular resolution, all neurons involved in perceiving and processing water flow information in larval zebrafish. We find a diverse array of neurons responding to forward flow, reverse flow, or both. Early in this pathway, in the lateral line ganglia, these are almost exclusively neurons responding to the simple presence of forward or reverse flow, but later processing includes neurons responding specifically to flow onset, representing the accumulated volume of flow during a stimulus, or encoding the speed of the flow. The neurons reporting on these more nuanced details are located across numerous brain regions, including some not previously implicated in water flow processing. A graph theory-based analysis of the brain-wide water flow network shows that a majority of this processing is dedicated to forward flow detection, and this is reinforced by our finding that details like flow velocity and the total volume of accumulated flow are only encoded for the simulated forward direction. The results represent the first brain-wide description of processing for this important modality, and provide a departure point for more detailed studies of the flow of information through this network.Significance statementIn aquatic animals, the lateral line is important for detecting water flow stimuli, but the brain networks that interpret this information remain mysterious. Here, we have imaged the activity of individual neurons across the entire brains of larval zebrafish, revealing all response types and their brain locations as water flow processing occurs. We find some neurons that respond to the simple presence of water flow, and others that are attuned to the flow’s direction, speed, duration, or the accumulated volume of water that has passed during the stimulus. With this information, we modeled the underlying network, describing a system that is nuanced in its processing of water flow simulating forward motion but rudimentary in processing flow in the reverse direction.

scholarly journals Competition between protons and substrate for binding to the major facilitator superfamily multidrug/H+ antiporter MdtM

2021 ◽  
Vol 2 ◽  
Author(s):  
Christopher J. Law
Keyword(s):  
Escherichia Coli ◽  
General Feature ◽  
Pathogenic Bacteria ◽  
Major Facilitator Superfamily ◽  
Electrochemical Gradient ◽  
Multidrug Efflux ◽  
Protein Fluorescence ◽  
Ph Values ◽  
Major Facilitator ◽  
Intrinsic Protein Fluorescence

Abstract Proton electrochemical gradient-driven multidrug efflux activity of representatives of the major facilitator superfamily (MFS) of secondary active transporters contributes to antimicrobial resistance of pathogenic bacteria. Integral to the mechanism of these transporters is a proposed competition between substrate and protons for the binding site of the protein. The current work investigated the competition between protons and antimicrobial substrate for binding to the Escherichia coli MFS multidrug/H+ antiporter MdtM by measuring the quench of intrinsic protein fluorescence upon titration of substrate tetraphenylphosphonium into a solution of purified MdtM over a range of pH values between pH 8.8 and 5.9. The results, which revealed that protons inhibit binding of substrate to MdtM in a competitive manner, are consistent with those reported in a study on the related MFS multidrug/H+ antiporter MdfA and provide further evidence that competition for binding between substrate and protons is a general feature of secondary multidrug efflux.

Shaking Table Tests on Multi-Leaf Stone Masonry Structures: Analysis of Stiffness Decay

2010 ◽  
Vol 133-134 ◽  
pp. 647-652 ◽  
Author(s):  
Nicola Mazzon ◽  
Cano M. Chavez ◽  
Maria Rosa Valluzzi ◽  
F. Casarin ◽  
Claudio Modena
Keyword(s):  
Dynamic Behaviour ◽  
Shaking Table ◽  
Stone Masonry ◽  
Mode Shapes ◽  
Masonry Structures ◽  
Shaking Table Tests ◽  
Grout Injection ◽  
Hydraulic Lime ◽  
Natural Hydraulic Lime ◽  
Before And After

The influence of the natural hydraulic lime-based grout on the dynamic behaviour of injected multi-leaf stone masonry elements is discussed in the paper. Shaking table experiments on two stone masonry buildings, tested before and after grout injection, have been performed. The paper focuses on the analysis of both the recorded accelerations and related displacements, at the bottom and at each further storey. This leads to evaluate the stiffness of the unstrengthened and injected structures. The input at increasing PGA allowed the stiffness decay to be studied, simulating a gradual damaging of the structures. These results were also interpreted in the light of both computed frequencies and mode shapes. Finally, the comparison among these results, obtained from all the models, allows to deepen the knowledge concerning the effects induced by the lime-based grout injection and on its capability to modify the dynamic behaviour, when intervening on a damaged (repairing) or on an undamaged (strengthening) structure.

scholarly journals Identification of C-terminal motifs responsible for transmission of inhibition by ATP of mammalian phosphofructokinase, and their contribution to other allosteric effects

2004 ◽  
Vol 377 (1) ◽  
pp. 77-84 ◽  
Author(s):  
Oscar H. MARTÍNEZ-COSTA ◽  
Carmen HERMIDA ◽  
Cristina SÁNCHEZ-MARTÍNEZ ◽  
Belén SANTAMARÍA ◽  
Juan J. ARAGÓN
Keyword(s):  
Point Mutations ◽  
Terminal Part ◽  
Protein Fluorescence ◽  
Binding Studies ◽  
C Terminus ◽  
Allosteric Effectors ◽  
Inhibitory Site ◽  
Intrinsic Protein Fluorescence ◽  
Strong Activator

Systematic deletions and point mutations in the C-terminal extension of mammalian PFK (phosphofructokinase) led us to identify Leu-767 and Glu-768 of the M-type isoform (PFK-M) as the motifs responsible for the role of this region in inhibition by MgATP. These amino acids are the only residues of the C-terminus that are conserved in all mammalian isoforms, and were found to have a similar function in the C-type isoenzyme. Both residues in PFK-C and Leu-767 in PFK-M were also observed to be critical for inhibition by citrate, which is synergistic with that by MgATP. Binding studies utilizing titration of intrinsic protein fluorescence indicated that the C-terminal part of the enzyme participates in the signal transduction route from the MgATP inhibitory site to the catalytic site, but does not contribute to the binding of this inhibitor, whereas it is essential for the binding of citrate. Mutations of the identified structural motifs did not alter either the action of other allosteric effectors that also interact with MgATP, such as the inhibitor phosphoenolpyruvate and the strong activator fructose 2,6-bisphosphate, or the co-operative effect of fructose 6-phosphate. The latter data provide evidence that activation by fructose 2,6-bisphosphate and fructose 6-phosphate co-operativity are not linked to the same allosteric transition as that mediating inhibition by MgATP.

Tyrosine Biosynthesis in Sorghum bicolor: Characteristics of Prephenate Aminotransferase

1986 ◽  
Vol 41 (1-2) ◽  
pp. 79-86 ◽  
Author(s):  
Daniel L. Siehl ◽  
James A. Connelly ◽  
Eric E. Conn
Keyword(s):  
Reaction Rate ◽  
Deae Cellulose ◽  
Forward Direction ◽  
Reverse Direction ◽  
Aminotransferase Activity ◽  
Forward Reaction ◽  
High Affinity ◽  
Sorghum Plant ◽  
Arogenate Dehydrogenase ◽  
Tyrosine Biosynthesis

Abstract A stable activity which transfers the amino group from glutamate to prephenate was extracted from 4-day old etiolated shoots of sorghum. The activity was retained on DEAE cellulose and eluted as a single peak. Prephenate aminotransferase co-eluted with a very abundant α-ketoglutarate: aspartate aminotransferase, but heating at 70 °C resulted in loss of α-ketoglutarate: aspar­tate activity with nearly full retention of prephenate: glutamate aminotransferase activity. The heated enzyme displayed high affinity and specificity for prephenate. Among 7 donors tested, only glutamate, and aspartate at less than 20% the rate with glutamate, supported prephenate aminotransferase activity. In the reverse direction, a reaction rate comparable to that in the forward direction was unchanged as the concentration of α-ketoglutarate was reduced from 1.0 to 0.09 mᴍ. The apparent Km for arogenate was 0.8 mᴍ. The forward reaction was unaffected by the inclusion of tyrosine, phenylalanine or tryptophan. Together with the discovery of arogenate dehydrogenase in sorghum [3], these data indicate that, in the sorghum plant, tyrosine derives from prephenate by transamination and aromatization. rather than the reverse sequence.

Sign in / Sign up

Export Citation Format

Share Document