Kinetics of Irreversible Inhibition of Creatine Kinase during Modification by o-Phthaldehyde

1994 ◽  
Vol 48 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Ze-Feng Wang ◽  
Yi-Kang Xu ◽  
Hai-Meng Zhou
Keyword(s):  
Creatine Kinase ◽  
Irreversible Inhibition ◽  
Kinetics Of

Inactivation and conformational changes of creatine kinase at low concentrations of hexafluoroisopropanol solutions

2003 ◽  
Vol 81 (5) ◽  
pp. 327-333 ◽  
Author(s):  
Xiao-Yun Wang ◽  
Fan-Guo Meng ◽  
Hai-Meng Zhou
Keyword(s):  
Creatine Kinase ◽  
Enzyme Activity ◽  
Conformational Changes ◽  
Fluorescence Spectra ◽  
Cd Spectra ◽  
Irreversible Inhibition ◽  
Low Concentrations ◽  
Flexible Region ◽  
Far Ultraviolet ◽  
Kinetics Of

Using the methods of far-ultraviolet circular dichroism (CD) spectra, fluorescence spectra, and enzyme activity assays, the inactivation and conformational changes of creatine kinase (CK) induced by 1,1,1,3,3,3-hexafluoro-2-propanol (hexafluoroisopropanol (HFIP)) of different concentrations were investigated. To avoid the aggregation of CK that occurs with high HFIP, concentrations of 0%–5% HFIP were used in this study. The CD spectra showed that HFIP concentrations above 2.5% strongly induced the formation of secondary structures of CK. No marked conformational changes were observed at low concentrations of HFIP (0%–2.5%). After incubation with 0.2% HFIP for 10 min, CK lost most of its activity. The kinetic theory of the substrate reaction during irreversible inhibition of enzyme activity described previously by Tsou was applied to study the kinetics of CK inactivation during denaturation by HFIP. The inactivation rate constants for the free enzyme and the substrate–enzyme complex were determined by Tsou's method. The results suggested that low concentrations of HFIP had a high potential to induce helices of protein and that the active site of the enzyme was situated in a limited and flexible region of the enzyme molecule that was more susceptible to the denaturant than was the protein as a whole.Key words: creatine kinase, inactivation, conformation, kinetics, hexafluoroisopropanol.

scholarly journals Activation of sea-urchin sperm motility is accompanied by an increase in the creatine kinase exchange flux

1997 ◽  
Vol 325 (2) ◽  
pp. 411-416 ◽  
Author(s):  
Ferdi A. van DORSTEN ◽  
Markus WYSS ◽  
Theo WALLIMANN ◽  
Klaas NICOLAY
Keyword(s):  
Creatine Kinase ◽  
Sea Urchin ◽  
Magnetization Transfer ◽  
Order Rate Constant ◽  
Phosphate Transport ◽  
High Energy ◽  
Sperm Activation ◽  
Kinetics Of ◽  
Sea Urchin Sperm ◽  
Exchange Flux

The kinetics of the creatine kinase (CK) reaction were studied in suspensions of quiescent and active, intact sea-urchin spermatozoa in artificial seawater, using 31P-NMR magnetization transfer. In inactive sperm, no CK-mediated exchange flux was detected, whereas in activated motile sperm, the forward pseudo-first-order rate constant was 0.13±0.04 s-1 at 10 °C, corresponding to a steady-state CK flux of 3.1±0.5 mM·s-1. Intracellular pH shifted from 6.6±0.1 to 7.6±0.1 upon activation. The phosphocreatine (PCr)/ATP and PCr/Pi ratios were only marginally reduced in activated sperm, whereas the estimated cytosolic free ADP concentration increased remarkably from 9 μM in quiescent, to 114 μM in activated spermatozoa. The elevation of CK flux upon sperm activation is discussed in the light of the proposition that in sea-urchin spermatozoa, which are fuelled entirely by oxidative phosphorylation, high-energy phosphate transport is mediated by a ‘CK/PCr shuttle‘.

Kinetics of the creatine kinase reaction in neonatal rabbit heart: an empirical analysis of the rate equation

Biochemistry ◽  
1991 ◽  
Vol 30 (10) ◽  
pp. 2585-2593 ◽  
Author(s):  
John J. McAuliffe ◽  
Stanton B. Perry ◽  
Elwood E. Brooks ◽  
Joanne S. Ingwall
Keyword(s):  
Creatine Kinase ◽  
Empirical Analysis ◽  
Rate Equation ◽  
Rabbit Heart ◽  
Kinase Reaction ◽  
Creatine Kinase Reaction ◽  
Kinetics Of

scholarly journals Kinetics of creatine uptake in the perfused mouse liver: a 31P-n.m.r. study of transgenic mice expressing creatine kinase (CKBB) in the liver

1994 ◽  
Vol 303 (2) ◽  
pp. 531-538 ◽  
Author(s):  
S Masson ◽  
B Quistorff
Keyword(s):  
Creatine Kinase ◽  
Transgenic Mice ◽  
Mouse Liver ◽  
Biochemical Analysis ◽  
Perfused Liver ◽  
Creatine Uptake ◽  
Total Creatine ◽  
Kinetics Of ◽  
Dependent Mechanism

Transport of creatine in the mouse liver has been investigated in vivo and in the perfused organ. Experiments were carried out with transgenic mice expressing creatine kinase in the liver (brain isoenzyme CKBB; EC 7.2.3.2.) [Koretsky, Brosnan, Chen, Chen and Van Dyke (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 3112-3116] and in the corresponding control mice. The animals were fed a regular chow with or without the addition of 10% creatine (w/w) for 5 days. The kinetics of creatine uptake was measured in the perfused liver by 31P-n.m.r. spectroscopy and biochemical analysis following infusion of creatine at concentrations ranging over 0-15 mM and at an extracellular pH of either 7.40 or 6.40. The results suggest that creatine is actively transported by a pH-dependent mechanism obeying a saturable Michaelis-Menten type of kinetics (Km = 0.80 +/- 0.18 and 5.12 +/- 2.40 mM; Vmax. = 0.57 +/- 0.04 and 1.72 +/- 0.32 mumol.g of liver-1.min-1 at pH 7.40 and 6.40 respectively). Creatine export was evaluated in the perfused liver preloaded with creatine and the results show that less than 2.5 and 5% of the total creatine pool is exported to the perfusate during 80 min of perfusion at pH 7.40 and 6.40 respectively. Taken together, these results seem to explain the observation that creatine accumulates in the mouse liver only when blood creatine is raised by creatine feeding.

Elimination kinetics of myoglobin and creatine kinase in rhabdomyolysis: Implications for follow-up

2002 ◽  
Vol 30 (10) ◽  
pp. 2212-2215 ◽  
Author(s):  
Helena Lappalainen ◽  
Eero Tiula ◽  
Lasse Uotila ◽  
Matti Mänttäri
Keyword(s):  
Creatine Kinase ◽  
Elimination Kinetics ◽  
Kinetics Of

Structural Analysis and Inhibitory Kinetics of Brain Type Creatine Kinase by Sodium Dodecyl Sulfate

2008 ◽  
Vol 160 (3) ◽  
pp. 831-842 ◽  
Author(s):  
Zhi-Rong Lü ◽  
Sang Ho Oh ◽  
Shan-Shan Zhou ◽  
He-Chang Zou ◽  
Daeui Park ◽  
...  
Keyword(s):  
Creatine Kinase ◽  
Structural Analysis ◽  
Sodium Dodecyl Sulfate ◽  
Sodium Dodecyl ◽  
Dodecyl Sulfate ◽  
Kinetics Of
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