We have successfully expressed and purified the three full-length human nonmuscle myosin II (NM II) proteins with their light chains, using the baculovirus/Sf9 system. The enzymatic and structural properties of the three paralogs were characterized. Although each NM II is capable of forming bipolar filaments, those formed by IIC tend to contain fewer constituent molecules than those of IIA and IIB. All paralogs adopt the compact conformation in the presence of ATP. Phosphorylation of the regulatory light chain leads to assembly into filaments, which bind to actin in the presence of ATP. The nature of interactions with actin filaments is shown with different paralogs exhibiting different actin binding behaviors under equivalent conditions. The data show that although NM IIA and IIB form filaments with similar properties, NM IIC forms filaments that are less well suited to roles such as tension maintenance within the cell. We are now extending these studies to include the use of mutant NM IIs that have been shown to play a causative role in human syndromes.

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Budget End
Support Year
24
Fiscal Year
2014
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U.S. National Heart Lung and Blood Inst
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