Stability of creatine kinase-3 in lyophilized materials.

1982 ◽  
Vol 28 (1) ◽  
pp. 41-44 ◽  
Author(s):  
V S Whitner ◽  
L G Morin ◽  
S S McKneally ◽  
E J Sampson
Keyword(s):  
Creatine Kinase ◽  
Kinase Activity ◽  
Arrhenius Plot ◽  
Gradient Elution ◽  
Creatine Kinase Activity ◽  
Chromatographic Technique ◽  
First Order ◽  
The Stability ◽  
Sulfhydryl Compounds ◽  
Stable Matrix

Abstract We examined the stability of creatine kinase (EC 2.7.3.2) isoenzyme-3 (CK-3) in lyophilized bovine albumin matrices in the presence and absence of various sulfhydryl compounds and ADP. We initially purified CK-3 from human myocardium and skeletal muscle by the batch-chromatographic technique and by gradient elution column chromatography to specific activities of 293 and 93 kU/g, respectively. To assess stability, we subjected the lyophilized materials to storage studies at 4, 25, 37, 42, 56, and 65 degrees C and compared first-order rate constants for the decay of creatine kinase activity at 42 degrees C. Our most stable matrix contained, per liter, 2 mmol of ADP and 10 mmol of N-acetylcysteine, and had an extrapolated first-order half-life (Arrhenius plot) at -20 degrees C of approximately 60000 years.

Effect of light and temperature on the stability of creatine kinase in human sera and controls.

1979 ◽  
Vol 25 (4) ◽  
pp. 625-628 ◽  
Author(s):  
B Perry ◽  
B Doumas ◽  
B Jendrzejczak
Keyword(s):  
Creatine Kinase ◽  
Room Temperature ◽  
Kinase Activity ◽  
Thermal Inactivation ◽  
Creatine Kinase Activity ◽  
Oxidative Process ◽  
Average Increase ◽  
Human Sera ◽  
The Stability ◽  
Effect Of Light

Abstract Commercial control preparations were exposed to light at room temperature for as long as 24 h. Decreases in creatine kinase activity in seven of the nine controls ranged from 28.8 to 83.3%. When these controls were protected from light and stored at various temperatures, the decrease in activity was either eliminated or substantially reduced, indicating that the decreased activity in the presence of light was not due entirely to thermal inactivation. In the absence of oxygen, light had little effect on creatine kinase suggesting that light inactivation is a light-catalyzed oxidative process. The decreased activity observed when some of the controls were exposed to light was not completely restored by incubating with dithiothreitol before analysis. However, increases in creatine kinase activity ranging from about 37 to 176% were observed when freshly prepared controls were pre-incubated with dithiothreitol. Neither light nor dithiothreitol had any effect on the creatine kinase activity in two of the nine controls. When human sera were exposed to light for 24 h, the largest decrease in activity was about 15%. Incubation of fresh human sera with dithiothreitol before analysis caused an average increase in activity of approximately 10%.

Separation of serum creatine kinase isoenzymes by ion-exchange column chromatography.

1977 ◽  
Vol 23 (3) ◽  
pp. 504-510 ◽  
Author(s):  
B Klein ◽  
J A Foreman ◽  
C L Jeunelot ◽  
J E Sheehan
Keyword(s):  
Creatine Kinase ◽  
Kinase Activity ◽  
Total Activity ◽  
Gradient Elution ◽  
Creatine Kinase Activity ◽  
Serum Creatine Kinase ◽  
Present Procedure ◽  
Analytical Recovery ◽  
Ion Exchange Column ◽  
Salt Gradient

Abstract We describe a practical, technically convenient DEAE-Sephadex chromatographic column and a three-fraction salt-gradient elution procedure, with which serum creatine kinase activity is rapidly and quantitatively separated into the isoenzyme components with excellent analytical recovery of the total activity applied. The homogeneity of the isoenzyme fractions was demonstrated by electrophoresis and rechromatography. Sera assessed by the Mercer fractionation [Clin. Chem. 20, 36 (1974)] and the present procedure showed excellent agreement.

Atypical increase in serum creatine kinase activity in hospital patients.

1976 ◽  
Vol 22 (1) ◽  
pp. 87-91 ◽  
Author(s):  
S M Sax ◽  
J J Moore ◽  
J L Giegel ◽  
M Welsh
Keyword(s):  
Creatine Kinase ◽  
Ion Exchange ◽  
Kinase Activity ◽  
Electrophoretic Analysis ◽  
Creatine Kinase Activity ◽  
Chromatographic Technique ◽  
Electrophoretic Method ◽  
Hospital Patients ◽  
Total Creatine Kinase Activity ◽  
Total Creatine

Abstract We use an ion-exchange column-chromatographic technique for separating creatine kinase isoenzymes in serum, and occasionally observe what appears to be sustained increase in the MB fraction. Most patients whose sera show such behavior have myocardial disease, but not necessarily a recent myocardial infarction. Electrophoretic analysis of a small sampling of such sera revealed that the apparent MB migrates atypically, appearing distinctly between isoezymes MB and MM. In another electrophoretic system, the peak might easily be mistaken for MM. This unusual isoenzyme does not appear to be "macro" creatine kinase. In laboratories that use the ion-exchange technique, the possibility of a falsely positive MB value should be considered in subjects who show persistent increases together with normal or nearly normal values for total creatine kinase activity. A suitable electrophoretic method that clearly demonstrates this unusual isoenzyme should be used in such cases, for confirmation.

Contractile economy and aerobic recovery metabolism in skeletal muscle adapted to creatine depletion

1994 ◽  
Vol 267 (1) ◽  
pp. C127-C137 ◽  
Author(s):  
T. S. Moerland ◽  
M. J. Kushmerick
Keyword(s):  
Creatine Kinase ◽  
Rate Constants ◽  
Kinase Activity ◽  
Citrate Synthase ◽  
Creatine Kinase Activity ◽  
Cellular Respiration ◽  
First Order ◽  
Myosin Isoenzymes ◽  
Total Creatine ◽  
Rate Limiting

Mice were treated for 7-12 wk with the creatine analogue beta-guanidinopropionic acid (beta-GPA). Treatment reduced total creatine to approximately 5% of control values in soleus (SOL) and extensor digitorum longus (EDL) muscles. In both muscles from treated mice, phosphorylated beta-GPA accumulated and resting [ATP] decreased by approximately 50%. Relative to controls, cytochrome oxidase and citrate synthase activities increased significantly in EDL from treated mice, but not in SOL; creatine kinase activity decreased significantly in SOL, but not in EDL. Measurements of poststimulation energy metabolism show that the energy cost to maintain tension in SOL and EDL from treated mice was approximately 50% of that in control muscle. Relative to controls, first-order rate constants of poststimulation O2 demand were 2- and 3.6-fold greater in SOL and EDL, respectively, from treated mice. Increased economy of SOL and EDL from treated mice is consistent with previously reported changes in myosin isoenzymes. Increases in rate constants of O2 utilization in creatine-depleted muscle are inconsistent with the hypothesis that cytoplasmic or mitochondrial creatine kinase is rate limiting for cellular respiration.

Distribution of serum creatine kinase activity in young healthy persons

1999 ◽  
Vol 279 (1-2) ◽  
pp. 107-115 ◽  
Author(s):  
Eli I. Lev ◽  
Ilan Tur-Kaspa ◽  
Isaac Ashkenazy ◽  
Anat Reiner ◽  
David Faraggi ◽  
...  
Keyword(s):  
Creatine Kinase ◽  
Kinase Activity ◽  
Creatine Kinase Activity ◽  
Serum Creatine Kinase ◽  
Healthy Persons

scholarly journals Ability of adenovirus 5 E1A proteins to suppress differentiation of BC3H1 myoblasts correlates with their binding to a 300 kDa cellular protein.

1992 ◽  
Vol 3 (10) ◽  
pp. 1107-1115 ◽  
Author(s):  
J S Mymryk ◽  
R W Lee ◽  
S T Bayley
Keyword(s):  
Creatine Kinase ◽  
Kinase Activity ◽  
Cellular Protein ◽  
Creatine Kinase Activity ◽  
Cellular Proteins ◽  
Deletion Mutants ◽  
Transcriptional Enhancers ◽  
Actin Mrna ◽  
Alpha Actin ◽  
Adenovirus 5

We have used deletion mutants to define the regions in Ad5 E1A proteins necessary to suppress differentiation of mouse BC3H1 myoblasts. We examined the differentiation of cells infected at a low multiplicity with viruses containing the E1A deletions and constructed so as to produce only the smaller of the two major E1A proteins. Only four of the mutant viruses containing deletions within the N-terminal 69 residues failed to suppress differentiation as judged by changes in morphology and in levels of muscle-specific alpha-actin mRNA and creatine kinase activity. The results were confirmed by analyses of lines of cells stably transfected with representative E1A mutants. The mouse cellular proteins to which mutant E1A proteins bound were identified by immunoprecipitating E1A proteins specifically from infected BC3H1 cells and by analyzing the precipitates on denaturing gels. Bands of proteins of 300, 130, 107, 105 (the retinoblastoma product), and 60 kDa (cyclin A) were distinguished. Failure to suppress differentiation correlated with loss of binding to the 300-kDa protein but not to any of the others. The regions of E1A defined in this way have been shown to be required for several other activities, including enhancer repression and transformation. One function of the 300-kDa protein appears to be to facilitate the action of transcriptional enhancers of differentiation-specific genes.

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