Effect of incubation in human plasma on electrophoretic mobility of brain-type creatine phosphokinase

doi:10.1016/0009-8981(76)90171-6

Clinica Chimica Acta

Volume 73, Issue 2, 1 December 1976, Pages 257-265

https://doi.org/10.1016/0009-8981(76)90171-6 Get rights and content

Abstract

1.
1. Electrophoretic mobility of brain-type creatine phosphokinase (CPK) from rabbit, rat and human decreased after incubation in human plasma for 3 h or 17 h but did not change after incubation in isotonic saline solution. The presence of the reducing agent mercaptoethanol during incubation in human plasma tended to reduce the rate of change of electrophoretic mobility of BB-CPK.
2.
2. The activity of brain CPK was almost completely lost after incubation for 17 h in human plasma. The addition of mercaptoethanol after incubation of brain CPK in human plasma could only partially restore the enzyme activity.
3.
3. The decrease in electrophoretic mobility of rat brain CPK occurred after incubation in an ultra-filtrate of human plasma which contained molecules whose molecular weight did not exceed 1000.
4.
4. Gel chromatography indicated that incubation of brain CPK in human plasma did not markedly change the molecular size of the enzyme.

References (20)

K.J. Van der Veen et al.
Clin. Chim. Acta
(1966)
J. Acheson et al.
Lancet
(1965)
A. Cao et al.
Clin. Chim. Acta
(1969)
H. Dubo et al.
Lancet
(1967)
A. Konttinen et al.
Am. J. Cardiol.
(1972)
T.D. Trainer et al.
Clin. Chim. Acta
(1968)
H.J. Keutel et al.
Arch. Biochem. Biophys.
(1972)
E. Jockers-Wretou et al.
Clin. Chim. Acta
(1975)
H.Y. Meltzer
Arch. Gen. Psychiatr.
(1969)
S.B. Rosalki
Nature
(1965)

There are more references available in the full text version of this article.

Cited by (24)

Evaluation of an immunoradiometric assay specific for the CK-MB isoenzyme for detection of acute myocardial infarction
1984, The American Journal of Cardiology
Clinical evaluation of patient's symptoms, electrocardiographic changes and increased serum enzyme levels, specifically creatine kinase (CK)-MB by electrophoresis, are established as the primary diagnostic indicators for myocardial infarction (Ml). Two hundred fifteen patients were evaluated in this study. Of these patients, 102 were admitted to the coronary care unit and 113 were admitted to the emergency room and screened for possible Ml. The immunoradiometric assay used in this study was a double antibody “sandwich” technique, which utilizes antibody to the M and B monomers of the CK isoenzymes. This assay is specific for the CK-MB isoenzyme, which is present in increased levels in MI. The intraassay coefficients of variation for 30 samples were 11.7% (mean 4.1 equivalent units [EU]/liter) and 8.4% (mean 15.4 EU/liter) and the interassay coefficients of variation for 30 samples were 11.1 % (mean 2.6 EU/liter) and 8.1 % (mean 13.6 EU/liter). The diagnostic sensitivity, specificity and accuracy in this study was 100%, respectively. The CK-MB by the immunoradiometric assay was found to be significantly more accurate than electrophoresis and, therefore, a reliable and also technically simpler replacement for electrophoresis.
An assay for creatine kinase conversion factor
1982, Clinical Biochemistry
The M subunits of creatine kinase (CK) isoenzymes, MM and MB, are subject to a postsynthetic modification in serum. As a result of this modification three sub-bands of MM can be identified by agarose electrophoresis, called MM₃, MM₂ and MM₁ in the order of increasing electrophoretic mobility towards the anode. An assay has been developed for the protein, called creatine kinase conversion factor, which catalyses the reaction leading to the sub-bands. The assay is based on the rate of conversion of MM₃ to MM₂ and MM₁. The first order rate constant for the conversion, designated as k_c, for sera of 24 apparently healthy adults ranges from 0.0221 to 0.0740 hr⁻¹ with a mean of 0.0392 hr⁻¹ under the conditions of the assay. Temperature and pH dependence of k_c have been determined.
Studies on the change of electrophoretic mobility and the decay of catalytic activity of brain-type creatine kinase isoenzyme (CK-BB) following incubation at 37 °C
1981, Clinica Chimica Acta
Incubation of CK-BB in serum (1:3, v/v) at 37°C for 3 h caused a change of its electrophoretic mobility and decay of its catalytic activity. Similar effects were observed following incubation in water. Incubation in saline somehow preserved the electrophoretic mobility but not the catalytic activity. No effect was noted when incubated at 4°C for 3 h. Further study on the rate of decay revealed that the decay in albumin solution (1:50, v/v) is quite similar to that in serum. More dramatic decay was noted when incubated in water and less when incubated in saline. It was further shown that the higher the incubation temperature (4°C, 25°C or 37°C) the faster the decay. The rate of decay of CK-MM was much slower in all conditions of incubation. Determination of isoenzyme activities by means of an immunoprecipitation method again demonstrated that CK-BB lost a great deal of its catalytic activity following incubation at 37°C for 1 h, and hence falsification of the isoenzyme pattern.
CREATINE KINASE MB IN ACUTE BRAIN INJURY
1978, The Lancet
HYPERTENSION, CONVULSIONS, AND CEREBRAL HÆMORRHAGE IN SICKLE-CELL ANÆMIA PATIENTS AFTER BLOOD-TRANSFUSIONS
1978, The Lancet
Effects of β-mercaptoethanol and chelating agents on the stability and activation of creatine kinase in serum
1978, Clinica Chimica Acta
We investigated the effects of sulfhydryl compounds on the stability of creatine kinase (CK) in unfrozen human serum and found that both β-mercaptoethanol and N-acetylcysteine led to accelerated loss of the endogenous serum enzyme activity. This is in contrast to the results of others who have either studied the stability of exogenous enzyme added to human serum or studied endogenous enzyme in frozen serum. The addition of cation chelators to serum markedly improved the stability of the endogenous CK activity. The enhanced stability was independent of chelation of calcium, iron, manganese, copper, or zinc. In addition, cation chelators caused a 16% increase in the CK activity of fresh samples. This latter effect was independent of the activation of CK by BME and could be accounted for by chelation of calcium ions during the assay. The data suggest that addition of cation chelators prior to storage may be useful in enhancing the stability of CK in serum whereas sulfhydryl compounds should be added prior to assay rather than prior to storage.

View all citing articles on Scopus

View full text

Effect of incubation in human plasma on electrophoretic mobility of brain-type creatine phosphokinase

Abstract

Clin. Chim. Acta

Lancet

Clin. Chim. Acta

Lancet

Am. J. Cardiol.

Clin. Chim. Acta

Arch. Biochem. Biophys.

Clin. Chim. Acta

Arch. Gen. Psychiatr.

Nature