The localization of mitochondrial creatine kinase, and its use for the determination of the sidedness of submitochondrial particles
| Autor: | E.M. Wit-Peeters, H.R. Scholte, P.J. Weijers |
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| Rok vydání: | 1973 |
| Předmět: |
Detergents
Guinea Pigs Biophysics Adenylate kinase Digitonin Mitochondrion Biochemistry Glycols chemistry.chemical_compound Animals Inner membrane Ultrasonics Aspartate Aminotransferases Submitochondrial particle Inner mitochondrial membrane Creatine Kinase Membranes biology Myocardium Phosphotransferases Cell Biology Adenosine Monophosphate Mitochondria Muscle Rats Microscopy Electron Membrane chemistry biology.protein Creatine kinase Polyethylenes Ethers |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Biomembranes. 291:764-773 |
| ISSN: | 0005-2736 |
| Popis: | 1. 1. Creatine kinase (EC 2.7.3.2) is extracted from heart mitochondria by suspension in salt-containing media. 2. 2. Digitonin treatment of isolated heart mitochondria causes rupture of the outer membranes and release of adenylate kinase (EC 2.7.4.3), while creatine kinase remains bound to the mitochondrial membranes. Although the outer membranes are ruptured by the action of digitonin, they can not be obtained in reasonable yield by differential centrifugation, indicating that most of the ruptured outer membranes are still connected to the inner membranes, which could be confirmed by electron microscopy. 3. 3. The addition of the detergent Lubrol WX to intact heart mitochondria has no effect upon the activity of the creatine kinase, indicating that this enzyme is not latent. 4. 4. In submitochondrial particles, obtained by ultrasonic treatment of heart mitochondria and removal of intact mitochondria by centrifugation, creatine kinase becomes partially latent. This indicates that in some of the submitochondrial particles the enzyme is localized inside the enveloping inner membrane fragment. Evidence is also presented for the existence of submitochondrial particles with the same orientation of the enveloping membrane as intact mitochondria, as indicated by the latency of aspartate aminotransferase (EC 2.6.1.1), a matrix enzyme that is firmly bound to the inside of the inner membrane. 5. 5. From Paragraphs 2–4 it is concluded that mitochondrial creatine kinase is bound to the outside of the inner mitochondrial membrane, in contrast to mitochondrial adenylate kinase, which is not bound to either of the mitochondrial membranes. |
| Databáze: | OpenAIRE |
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