title: Four things to know about myosin light chains as reporters for non-muscle myosin-2 dynamics in live cells.
abstract: The interplay between non-muscle myosins-2 and filamentous actin results in cytoplasmic contractility which is essential for eukaryotic life. Concomitantly, there is tremendous interest in elucidating the physiological function and temporal localization of non-muscle myosin-2 in cells. A commonly used method to study the function and localization of non-muscle myosin-2 is to overexpress a fluorescent protein (FP)-tagged version of the regulatory light chain (RLC) which binds to the myosin-2 heavy chain by mass action. Caveats about this approach include findings from recent studies indicating that the RLC does not bind exclusively to the non-muscle myosin-2 heavy chain. Rather, it can also associate with the myosin heavy chains of several other classes as well as other targets than myosin. In addition, the presence of the FP moiety may compromise myosin's enzymatic and mechanical performance. This and other factors to be discussed in this commentary raise questions about the possible complications in using FP-RLC as a marker for the dynamic localization and regulatory aspects of non-muscle myosin-2 motor functions in cell biological experiments.
authors.affiliation: Laboratory of Molecular Physiology, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland.
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Heissler
authors.forename: James R
authors.initials: JR
authors.lastname: Sellers
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D002465:Cell Movement; D003599:Cytoskeleton / Q000378:metabolism; D006801:Humans; D009119:Muscle Contraction / Q000502:physiology*; D018995:Myosin Heavy Chains / Q000378:metabolism*; D018994:Myosin Light Chains / Q000378:metabolism*; D010766:Phosphorylation
chemical_list: D000199:Actins; D018994:Myosin Light Chains; D018995:Myosin Heavy Chains
keywords: GFP; cell motility; myosin; phosphomimetics; regulatory light chain