scholarly journals Kinetic properties of cerebral pyruvate kinase

1974 ◽  
Vol 141 (1) ◽  
pp. 165-171 ◽  
Author(s):  
Peter C. Nicholas ◽  
Herman S. Bachelard
Keyword(s):  
Pyruvate Kinase ◽  
Enzymic Activity ◽  
Kinetic Properties ◽  
Muscle Enzyme ◽  
Mammalian Liver ◽  
Substrate Saturation ◽  
Enzyme Stimulation ◽  
High Concentrations ◽  
The Brain ◽  
Guinea Pig Brain

Partly purified guinea-pig brain pyruvate kinase is not activated by fructose 1,6-diphosphate and gives hyperbolic substrate-saturation curves with phosphoenolpyruvate. It is therefore different from the L-type pyruvate kinase of mammalian liver. Inhibition by MgATP2- was competitive for MgADP- but not for phosphoenolpyruvate, and the enzyme is therefore different from the M-type pyruvate kinase, which is said to be competitively inhibited by MgATP2- with respect to both substrates. The Ki(MgATP2-) value of approx. 8mm for the brain enzyme is higher than the values (about 2mm) reported for the muscle enzyme. Stimulation of enzymic activity was observed at low (1–2mm) concentrations of MgATP2-. Substrate kinetic constants were Km (MgADP-)=0.47mm, Km (phosphoenolpyruvate)=0.08mm. Free Mg2+ at very high concentrations (over 10mm) was inhibitory (Ki=20–32mm). Neither ADP3- nor 5′-AMP2- inhibited the activity. The brain enzyme was concluded to be different from both the M-type and the L-type of other mammalian organs such as muscle and liver.

scholarly journals Properties and reaction with iodoacetamide of adenosine 5′-triphosphate–creatinine phosphotransferase from human skeletal muscle. Further evidence about the role of the essential thiol group in relation to the mechanism of action

1970 ◽  
Vol 117 (3) ◽  
pp. 513-523 ◽  
Author(s):  
I. Kumudavalli ◽  
B. H. Moreland ◽  
D. C. Watts
Keyword(s):  
Skeletal Muscle ◽  
Transition State ◽  
Thiol Group ◽  
Equilibrium Mixture ◽  
Common Mechanism ◽  
Kinetic Properties ◽  
Rabbit Muscle ◽  
Muscle Enzyme ◽  
Similar Chemical ◽  
High Concentrations

1. The purification of creatine kinase from human and monkey skeletal muscle by horizontal electrophoresis on Sephadex blocks is described. 2. The purified enzymes are shown to have similar chemical and kinetic properties to the rabbit muscle enzyme and a common mechanism is inferred. 3. Iodoacetamide has a similar apparent second-order inhibition constant with the human and rabbit enzymes, but the inhibition does not go to completion with the former. This is even more marked with the monkey enzyme, which has more reactive thiol groups, but inhibition is only about 50%. 4. Single substrates have little effect on the inhibition by iodoacetamide, but with the primate enzymes, in contrast with the rabbit enzyme, high concentrations of ADP–Mg2+ plus creatine convert the essential thiol group from being pH-independent into one with a normal ionization. Low concentrations of ADP–Mg2+ plus creatine first enhance the rate of inactivation, but cause protection as the reaction proceeds. These results are interpreted to indicate an activation of the thiol group on the subunit to which the substrates bind and a co-operatively induced decrease in the activity of the thiol group on the other subunit which lacks substrates. 5. The effects of a substrate equilibrium mixture on the rate of inhibition are essentially those of ADP–Mg2+ plus creatine. 6. Since no substrate combination affords significant protection to the thiol group associated with the catalytic site to which the substrates are bound, it is concluded that any mechanism involving the thiol group in a direct participation in the transition-state complex of the catalytic reaction must be abandoned unless the transition state is only a small part of the time taken for one catalytic cycle.

Native Ubiquitin Structural Changes Resulting from Complexation with β-methylamino-L-alanine (BMAA)

2020 ◽  
Author(s):  
Katie Mae Wilson ◽  
Aurora Burkus-Matesevac ◽  
Samuel Maddox ◽  
Christopher Chouinard
Keyword(s):  
Structural Changes ◽  
Noncovalent Complex ◽  
Unfolded Protein ◽  
Protein Size ◽  
High Concentrations ◽  
The Unfolded Protein Response ◽  
Critical Binding ◽  
Protein Response ◽  
The Brain ◽  
Lateral Sclerosis

β-methylamino-L-alanine (BMAA) has been linked to the development of neurodegenerative (ND) symptoms following chronic environmental exposure through water and dietary sources. The brains of those affected by this condition, often referred to as amyotrophic lateral sclerosis-parkinsonism-dementia complex (ALS-PDC), have exhibited the presence of plaques and neurofibrillary tangles (NFTs) from protein aggregation. Although numerous studies have sought to better understand the correlation between BMAA exposure and onset of ND symptoms, no definitive link has been identified. One prevailing hypothesis is that BMAA acts a small molecule ligand, complexing with critical proteins in the brain and reducing their function. The objective of this research was to investigate the effects of BMAA exposure on the native structure of ubiquitin. We hypothesized that formation of a Ubiquitin+BMAA noncovalent complex would alter the protein’s structure and folding and ultimately affect the ubiquitinproteasome system (UPS) and the unfolded protein response (UPR). Ion mobility-mass spectrometry revealed that at sufficiently high concentrations BMAA did in fact form a noncovalent complex with ubiquitin, however similar complexes were identified for a range of additional amino acids. Collision induced unfolding (CIU) was used to interrogate the unfolding dynamics of native ubiquitin and these Ubq-amino acid complexes and it was determined that complexation with BMAA led to a significant alteration in native protein size and conformation, and this complex required considerably more energy to unfold. This indicates that the complex remains more stable under native conditions and this may indicate that BMAA has attached to a critical binding location.

Native Ubiquitin Structural Changes Resulting from Complexation with β-methylamino-L-alanine (BMAA)

2020 ◽  
Author(s):  
Katie Mae Wilson ◽  
Aurora Burkus-Matesevac ◽  
Samuel Maddox ◽  
Christopher Chouinard
Keyword(s):  
Structural Changes ◽  
Noncovalent Complex ◽  
Unfolded Protein ◽  
Protein Size ◽  
High Concentrations ◽  
The Unfolded Protein Response ◽  
Critical Binding ◽  
Protein Response ◽  
The Brain ◽  
Lateral Sclerosis

β-methylamino-L-alanine (BMAA) has been linked to the development of neurodegenerative (ND) symptoms following chronic environmental exposure through water and dietary sources. The brains of those affected by this condition, often referred to as amyotrophic lateral sclerosis-parkinsonism-dementia complex (ALS-PDC), have exhibited the presence of plaques and neurofibrillary tangles (NFTs) from protein aggregation. Although numerous studies have sought to better understand the correlation between BMAA exposure and onset of ND symptoms, no definitive link has been identified. One prevailing hypothesis is that BMAA acts a small molecule ligand, complexing with critical proteins in the brain and reducing their function. The objective of this research was to investigate the effects of BMAA exposure on the native structure of ubiquitin. We hypothesized that formation of a Ubiquitin+BMAA noncovalent complex would alter the protein’s structure and folding and ultimately affect the ubiquitinproteasome system (UPS) and the unfolded protein response (UPR). Ion mobility-mass spectrometry revealed that at sufficiently high concentrations BMAA did in fact form a noncovalent complex with ubiquitin, however similar complexes were identified for a range of additional amino acids. Collision induced unfolding (CIU) was used to interrogate the unfolding dynamics of native ubiquitin and these Ubq-amino acid complexes and it was determined that complexation with BMAA led to a significant alteration in native protein size and conformation, and this complex required considerably more energy to unfold. This indicates that the complex remains more stable under native conditions and this may indicate that BMAA has attached to a critical binding location.

Enzymic activity of the brain: microvessels vs. total forebrain homogenate

1977 ◽  
Vol 138 (3) ◽  
pp. 561-564 ◽  
Author(s):  
B.M. Djuricˇic´ ◽  
B.B. Mruˇlja
Keyword(s):  
Enzymic Activity ◽  
Brain Microvessels ◽  
The Brain

scholarly journals Ribonucleic acid stimulation of mammalian liver nuclear-envelope nucleoside triphosphatase. A possible enzymic marker for the nuclear envelope

1977 ◽  
Vol 162 (3) ◽  
pp. 671-679 ◽  
Author(s):  
P S Agutter ◽  
J R Harris ◽  
I Stevenson
Keyword(s):  
Nuclear Envelope ◽  
Specific Activity ◽  
Assay Medium ◽  
Exogenous Rna ◽  
Mammalian Liver ◽  
High Concentrations ◽  
Enzymic Marker ◽  
Stimulation Of ◽  
Nucleoside Triphosphatase

1. The specific activity of rat and pig liver nuclear-envelope nucleoside triphosphatase (EC 3.6.1.3) decreases when the system is depleted of RNA. The activity can be restored by adding high concentrations of yeast RNA to the assay medium. 2. Exogenous RNA also increases the activity of the enzyme in control envelopes (not RNA-depleted). The effect appears to be largely specific for poly(A) and poly(G); it is not stimulated by rRNA or tRNA preparations, ribonuclease-hydrolysed RNA, AMP, or double- or single-stranded DNA. 3. Inhibitors of the enzyme, in concentrations at which half-maximal inhibition of the enzyme is achieved, do not affect the percentage stimulation of the enzyme by yeast RNA. 4. The simulation is abolished by the inclusion of 150 mM-KCl or -NaCl in the assay medium, but not by increasing the assay pH to 8.5. 5. The results are discussed in the light of the possible role of the nucleoside triphosphatase in vivo in nucleo-cytoplasmic ribonucleoprotein translocation. 6. It is proposed that poly(G)-stimulated Mg2+-activated adenosine triphosphatase activity should be adopted as an enzymic marker for the nuclear envelope.

scholarly journals Use of 19F NMR Spectroscopy for Measurement of Cerebral Blood Flow: A Comparative Study Using Microspheres

1989 ◽  
Vol 9 (6) ◽  
pp. 886-891 ◽  
Author(s):  
David Barranco ◽  
Leslie N. Sutton ◽  
Sandra Florin ◽  
Joel Greenberg ◽  
Teresa Sinnwell ◽  
...  
Keyword(s):  
Blood Flow ◽  
Cerebral Blood Flow ◽  
Arterial Blood ◽  
Radioactive Microsphere ◽  
Low Concentrations ◽  
Cat Brain ◽  
Nmr Techniques ◽  
High Concentrations ◽  
Washout Curves ◽  
The Brain

19F NMR was used to determine washout curves of an inert, diffusible gas (CHF3) from the cat brain. The cerebral blood flow was estimated from a bi- or tri-phasic fit to the deconvoluted wash-out curve, using the Kety-Schmidt approach. Cerebral blood flow values determined by 19F NMR show the expected responsiveness to alterations in Paco2, but are approximately 28% lower than cerebral blood flow values determined simultaneously by radioactive microsphere techniques. High concentrations of CHF3 have little effect on intracranial pressure, mean arterial blood pressure or Paco2, but cause small changes in the blood flow to certain regions of the brain. We conclude that 19F NMR techniques utilizing low concentrations of CHF3 have potential for the noninvasive measurement of cerebral blood flow.

scholarly journals Effect of Handling on ATP Utilization of Cerebral Na,K-ATPase in rats with Trimethyltin-Induced Neurodegeneration.

2021 ◽  
Author(s):  
Barbora Kalocayova ◽  
Denisa Snurikova ◽  
Jana Vlkovicova ◽  
Veronika Navarova Stara ◽  
Dominika Michalikova ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Kinetic Properties ◽  
Potassium Ions ◽  
Catalytic Subunits ◽  
Key Enzyme ◽  
Male Wistar Rats ◽  
Frontal Cerebral Cortex ◽  
Α3 Subunit ◽  
The Brain ◽  
Sodium And Potassium

Abstract Previously it was shown that for reduction of anxiety and stress of experimental animals, preventive handling seems to be one of the most effective methods. The present study was oriented on Na,K-ATPase, a key enzyme for maintaining proper concentrations of intracellular sodium and potassium ions. Malfunction of this enzyme has an essential role in the development of neurodegenerative diseases. It is known that this enzyme requires approximately 50% of the energy available to the brain. Therefore in the present study utilization of the energy source ATP by Na,K-ATPase in the frontal cerebral cortex, using the method of enzyme kinetics was investigated. As a model of neurodegeneration treatment with Trimethyltin (TMT) was applied. Daily handling (10 min/day) of healthy rats and rats suffering neurodegeneration induced by administration of TMT in a dose of (7.5 mg/kg), at postnatal days 60-102 altered the expression of catalytic subunits of Na,K-ATPase as well as kinetic properties of this enzyme in frontal cerebral cortex of adult male Wistar rats. Everyday handling of rats, beside the previously published beneficial effect on spatial memory was accompanied by improwed maintenance of sodium homeeostasis in frontal cortex of brains. The key system responsible for this proces, the Na,K-ATPase was able to utilize better the energy substrate ATP. In rats with TMT-induced neurodegeneration handling promoted the expresion of α2 isoform of the enzyme which is typical for glial cells. In healthy rats the handling was followed by increased expression α3 subunit which is typical for neurons.

scholarly journals The representation of colored objects in macaque color patches

2017 ◽  
Author(s):  
Le Chang ◽  
Pinglei Bao ◽  
Doris Y. Tsao
Keyword(s):  
Object Recognition ◽  
Color Vision ◽  
The Other ◽  
Color Patch ◽  
Shape Information ◽  
Object Shape ◽  
Shape Invariant ◽  
High Concentrations ◽  
The Brain

AbstractAn important question about color vision is: how does the brain represent the color of an object? The recent discovery of “color patches” in macaque inferotemporal (IT) cortex, the part of brain responsible for object recognition, makes this problem experimentally tractable. Here we record neurons in three color patches, middle color patch CLC (central lateral color patch), and two anterior color patches ALC (anterior lateral color patch) and AMC (anterior medial color patch), while presenting images of objects systematically varied in hue. We found that all three patches contain high concentrations of hue-selective cells, and the three patches use distinct computational strategies to represent colored objects: while all three patches multiplex hue and shape information, shape-invariant hue information is much stronger in anterior color patches ALC/AMC than CLC; furthermore, hue and object shape specifically for primate faces/bodies are over-represented in AMC but not in the other two patches.

Kinetic properties of the muscular pyruvate kinase from the giant marine barnacle, Austromegabalanus psittacus (Molina, 1782) (Cirripedia, Balanomorpha)

Crustaceana ◽  
2005 ◽  
Vol 78 (10) ◽  
pp. 1203-1218
Author(s):  
Daniel López ◽  
Robert Simpfendörfer ◽  
Karin Oelckers ◽  
David Nash
Keyword(s):  
Pyruvate Kinase ◽  
Kinetic Properties
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