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A Preparative Method for the Isolation of Calponin from Molluscan Catch Muscle
Stanislav S Lazarev1, Ulyana V Shevchenko1, Vyacheslav A Dyachuk1
1Laboratory of Cell Biophysics, A.V. Zhirmunsky National Scientific Center of Marine Biology, Far Eastern Branch, Russian Academy of Sciences, 17 Palchevsky Str., 690041 Vladivostok, Russia.
International Journal of Molecular Sciences
|July 27, 2022
Summary
A new method efficiently isolates molluscan calponin using temperature-dependent actin interactions. This purified calponin modulates actomyosin Mg2+-ATPase activity, acting as both an inhibitor and inducer.
Area of Science:
- Biochemistry
- Muscle Physiology
- Protein Purification
Background:
- Calponin is a key regulatory protein in muscle contraction.
- Existing methods for calponin isolation have limitations in yield and purity.
Purpose of the Study:
- To develop an improved, preparative method for isolating functional molluscan calponin.
- To assess the functional activity of the isolated calponin on actomyosin ATPase activity.
Main Methods:
- Thin filaments were extracted from molluscan muscle.
- Calponin was isolated from actin via temperature-dependent ultracentrifugation at 2°C.
- Purification was achieved using cation-exchange chromatography.
Main Results:
- The new method yielded four-fold higher pure calponin compared to high-temperature extraction.
- Isolated calponin demonstrated both inhibition and induction of actomyosin Mg2+-ATPase activity.
- Calponin's functional effects were dependent on environmental conditions and the source of actin and myosin.
Conclusions:
- The developed method provides a highly efficient means to obtain pure, functional molluscan calponin.
- Calponin's ability to modulate actin conformation is crucial for its dual role in regulating ATPase activity.
- This purified calponin is suitable for further functional and structural studies.
