Purification of native myosin filaments from muscle

Biophysical Journal
C HidalgoR Craig

Abstract

Analysis of the structure and function of native thick (myosin-containing) filaments of muscle has been hampered in the past by the difficulty of obtaining a pure preparation. We have developed a simple method for purifying native myosin filaments from muscle filament suspensions. The method involves severing thin (actin-containing) filaments into short segments using a Ca(2+)-insensitive fragment of gelsolin, followed by differential centrifugation to purify the thick filaments. By gel electrophoresis, the purified thick filaments show myosin heavy and light chains together with nonmyosin thick filament components. Contamination with actin is below 3.5%. Electron microscopy demonstrates intact thick filaments, with helical cross-bridge order preserved, and essentially complete removal of thin filaments. The method has been developed for striated muscles but can also be used in a modified form to remove contaminating thin filaments from native smooth muscle myofibrils. Such preparations should be useful for thick filament structural and biochemical studies.

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Citations

Mar 5, 2003·Journal of Molecular Biology·Fa-Qing Zhao, Roger Craig
Feb 7, 2008·Proceedings of the National Academy of Sciences of the United States of America·Maria E ZoghbiRoger Craig
May 23, 2008·Molecular Biology of the Cell·Hyun Suk JungRoger Craig
Nov 4, 2008·Journal of Molecular Biology·Fa-Qing ZhaoJohn L Woodhead
Sep 9, 2008·Journal of Molecular Biology·Hyun Suk Jung, Roger Craig
Mar 28, 2006·Current Opinion in Structural Biology·Roger Craig, John L Woodhead
Nov 1, 2005·Biophysical Journal·Sergey V PlotnikovWilliam A Mohler
Aug 27, 2005·Nature·John L WoodheadRaúl Padrón
Jul 1, 2005·Physiological Reviews·Scott L Hooper, Jeffrey B Thuma
Jun 7, 2018·Acta Crystallographica. Section D, Structural Biology·Ieva DrulyteRebecca F Thompson

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