Isoelectric focusing and hybridisation experiments on creatine kinase (EC 2.7.3.2)

Wevers, Ron A.; Wolters, R.J.; Soons, J.B.J.

doi:10.1016/0009-8981(77)90316-3

Cited by 59 publications

(13 citation statements)

References 3 publications

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“…The influence of the modification in the CK M-chain is not restricted to the alteration in the u value. It has already been described that the isoelectric point changes as well [12]. We have found a third property of the enzyme that appears to be changed upon modification of the M-chain.…”

Section: Discussionsupporting

confidence: 62%

“…This explains why the modification cannot be demonstrated in buffer matrices that do not contain the modifying protein. The modification in the M-chain causes obvious changes in the Michaelis-Menten constants, the apparent activation energy and the isoelectric point [12] of the enzyme. All three parameters can be used as an index for the mean age of the CK M-chain in the blood.…”

Section: Discussionmentioning

confidence: 99%

“…They were redissolved in a 10 mmol/l Tris-HCl buffer, pH 7.4. The CK MM forms were separated from each other by column isoelectric focusing as described before [12]. Only the top fractions of the three CK MM activity peaks were used.…”

Section: Methodsmentioning

confidence: 99%

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Indices for the age of the creatine kinase M-chain in the blood

Wevers

Hagelauer

Stein

et al. 1985

Clinica Chimica Acta

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SummaryThe apparent activation energy of the CK reaction as well as the Michaelis-Menten constants and the isoelectric point of CK MM can be used as indices for the mean age of the CK M-chain in the blood in vivo and in vitro. Modifications in the CK M-chain take place in vivo in the blood and in vitro in a serum matrix. Gradual increases in the apparent activation energy are also observed both in vivo and in vitro. It is confirmed that the modification in the CK M-chain causes a rise in the apparent activation energy, u.A gradual increase in apparent activation energy, due to the ageing process of the CK M-chain, was observed after myocardial infarction. A significantly increased value for u was observed at the time that total CK activity already had returned to reference values. In spite of the normal CK value, the apparent activation energy still indicated that there had been myocardial damage. The Michaelis-Menten constants for creatine phosphate and ADP are also influenced significantly by the modification in the M-chain. While the apparent activation energy increases, the Michaelis constants decrease in the order MM,, MM,, MM,. The Michaelis-Menten constants for both ADP and CrP can be used as an index for the mean age of the enzyme in the blood.The Michaelis-Menten constants for CrP and ADP show significant variations with the measuring temperature for virtually all CK MM forms.

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Section: Discussionsupporting

confidence: 62%

Section: Discussionmentioning

confidence: 99%

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Indices for the age of the creatine kinase M-chain in the blood

Wevers

Hagelauer

Stein

et al. 1985

Clinica Chimica Acta

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“…The changes introduced in the M-chain do not affect the molecular mass measurably [2], nor the immunological properties [2]. In addition, the post-synthetic multiple forms of the enzyme do have enzymatic activity [3]. The minor alterations in the CK molecule, however, result in a significant decrease in isoelectric point as the modification process proceeds [3].…”

Section: Introductionmentioning

confidence: 99%

“…In addition, the post-synthetic multiple forms of the enzyme do have enzymatic activity [3]. The minor alterations in the CK molecule, however, result in a significant decrease in isoelectric point as the modification process proceeds [3]. The significance of this post-synthetic modification process is unknown.…”

Section: Introductionmentioning

confidence: 99%

Purification and determination of the modifying protein responsible for the post-synthetic modification of creatine kinase (EC 2.7.3.2) and enolase(EC 4.2.1.11)

Landeghem

Soons

Wever

et al. 1985

Clinica Chimica Acta

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SummaryThe purification of a serum protein, responsible for the postsynthetic modification of CK and enolase, is described. A purification of about 1300-fold could be reached after subsequent chromatography of human serum on DEAE cellulose and Sephacryl S-200 Superfine followed by affinity chromatography using antibodies against human serum albumin, C3 and C4 and against total human serum proteins. A recovery of 160% of modifying activity was found. The molecular mass and the isoelectric point of the modifying protein have been determined.It is concluded that the concentration of the modifying protein in human serum is < 210 mg/l.

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Heterogeneity of serum creatine kinase isoenzyme‐MM in acute myocardial infarction

Williams

Marshall

1989

Electrophoresis

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Isoelectric focusing of serum creatine kinase (CK;EC 2.7.3.2) reveals up to 14 CK-MM subbands following acute myocardial infarction (AMI). The "normal" subbands 1 (pI 6.91), 2 (pI 6.65) and 3 (pI 6.35) are faintly present in normal serum and the "abnormal" subbands c (pI 7.25), e (pI 6.85), g (pI 6.50), i (pI 6.28), j (pI 6.20) and k (pI 6.15) are prominently detected in sera with elevated CK. "Abnormal" subbands a (pI 7.55),b(pI7.35),d(pI7.05),f(pI6.72) and h(pI6.40) have only been detected in AMI. The "abnormal" subbands appear, and reach maximum intensity (together with CK-MM 1-3), 3-12 h after infarction, and become faint and anodally convert (as do CK-MM 1-3) within 36 h. Similar changes are detected by nonequilibrium pH gradient electrophoresis which combines CK-MM and CK-MB analysis. In vitro incubation of serum with 0.015 M 2-mercaptoethanol induces conversion of CK-MM 1, 2 and 3 to b and c, d and e, and f and g, respectively. Thus, the complexity of the patterns is explained by a secondary conversion of "normal" to "abnormal" subbands superimposed upon anodal conversion of CK-MM 1----3. The clinical significance of these findings is discussed.

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Isoelectric focusing and hybridisation experiments on creatine kinase (EC 2.7.3.2)

Cited by 59 publications

References 3 publications

Indices for the age of the creatine kinase M-chain in the blood

Indices for the age of the creatine kinase M-chain in the blood

Purification and determination of the modifying protein responsible for the post-synthetic modification of creatine kinase (EC 2.7.3.2) and enolase(EC 4.2.1.11)

Heterogeneity of serum creatine kinase isoenzyme‐MM in acute myocardial infarction

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