ArticleMolecular biology of the cell1995
A fluorescent protein biosensor of myosin II regulatory light chain phosphorylation reports a gradient of phosphorylated myosin II in migrating cells.
Article in Molecular biology of the cell, 1995. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 60 citations in OpenAlex.
- A Genetically Encoded Biosensor Strategy for Quantifying Non-muscle Myosin II Phosphorylation Dynamics in Living Cells and Organisms.Cell reports · 2018Article
- aPKCζ affects directed cell migration through the regulation of myosin light chain phosphorylation.Cell adhesion & migration · 2017Article
- Fluorescent protein biosensors applied to microphysiological systems.Experimental biology and medicine (Maywood, N.J.) · 2015Review
- Four things to know about myosin light chains as reporters for non-muscle myosin-2 dynamics in live cells.Cytoskeleton (Hoboken, N.J.) · 2015Article
- In vitro platforms for evaluating liver toxicity.Experimental biology and medicine (Maywood, N.J.) · 2014Review
- Studies on the interaction mechanism of pyrene derivatives with human tumor-related DNA.Molecules (Basel, Switzerland) · 2012Article
- Protrusion and actin assembly are coupled to the organization of lamellar contractile structures.Experimental cell research · 2010Article
- INSIGHTS INTO THE ROLES OF NON-MUSCLE MYOSIN IIA IN HUMAN KERATINOCYTE MIGRATION.Cellular and molecular bioengineering · 2009Article
- Visualizing and quantifying adhesive signals.Current opinion in cell biology · 2008Review
- Myosin phosphatase target subunit: Many roles in cell function.Biochemical and biophysical research communications · 2008Review
- Distinct roles of MLCK and ROCK in the regulation of membrane protrusions and focal adhesion dynamics during cell migration of fibroblasts.The Journal of cell biology · 2004Article
- Asymmetric distribution of myosin IIB in migrating endothelial cells is regulated by a rho-dependent kinase and contributes to tail retraction.Molecular biology of the cell · 2003Article
- Phosphorylation of myosin II regulatory light chain is necessary for migration of HeLa cells but not for localization of myosin II at the leading edge.The Biochemical journal · 2003Article
- A structural model for phosphorylation control of Dictyostelium myosin II thick filament assembly.The Journal of cell biology · 1999Article
- Keratocytes generate traction forces in two phases.Molecular biology of the cell · 1999Article
- The actin-based nanomachine at the leading edge of migrating cells.Biophysical journal · 1999Article
- Activation of myosin phosphatase targeting subunit by mitosis-specific phosphorylation.The Journal of cell biology · 1999Article
- Ca2+-independent myosin II phosphorylation and contraction in chicken embryo fibroblasts.The Journal of physiology · 1999Article
- Specific localization of serine 19 phosphorylated myosin II during cell locomotion and mitosis of cultured cells.The Journal of cell biology · 1998Article
- Differential localization of myosin and myosin phosphatase subunits in smooth muscle cells and migrating fibroblasts.Molecular biology of the cell · 1997Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Phosphorylation of the regulatory light chain by myosin light chain kinase (MLCK) regulates the motor activity of smooth muscle and nonmuscle myosin II. We have designed reagents to detect this phosphorylation event in living cells. A new fluorescent protein biosensor of myosin II regulatory light chain phosphorylation (FRLC-Rmyosin II) is described here. The biosensor depends upon energy transfer from fluorescein-labeled regulatory light chains to rhodamine-labeled essential and/or heavy chains. The energy transfer ratio increases by up to 26% when the regulatory light chain is phosphorylated by MLCK. The majority of the change in energy transfer is from regulatory light chain phosphorylation by MLCK (versus phosphorylation by protein kinase C). Folding/unfolding, filament assembly, and actin binding do not have a large effect on the energy transfer ratio. FRLC-Rmyosin II has been microinjected into living cells, where it incorporates into stress fibers and transverse fibers. Treatment of fibroblasts containing FRLC-Rmyosin II with the kinase inhibitor staurosporine produced a lower ratio of rhodamine/fluorescein emission, which corresponds to a lower level of myosin II regulatory light chain phosphorylation. Locomoting fibroblasts containing FRLC-Rmyosin II showed a gradient of myosin II phosphorylation that was lowest near the leading edge and highest in the tail region of these cells, which correlates with previously observed gradients of free calcium and calmodulin activation. Maximal myosin II motor force in the tail may contribute to help cells maintain their polarized shape, retract the tail as the cell moves forward, and deliver disassembled subunits to the leading edge for incorporation into new fibers.
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