Product Uses Include
Myosin II protein has been purified from rabbit skeletal muscle. The full length myosin II protein has been purified with its essential light chains (ELC) and regulatory light chains (RLC), see Figure 1 and 2. Myosin II has been determined to be biologically active in an F-actin activated ATPase assay. Rabbit skeletal muscle myosin II is not recommended for use in motility assays. Rabbit myosin II protein is supplied as a white lyophilized powder. When reconstituted in nanopure water, the protein will be in the following buffer: 25 mM PIPES-NaOH pH 7.0, 1.25 M KCl, 2.5% sucrose and 0.5% dextran. The lyophilized protein is stable at 4°C desiccated (<10% humidity) for 1 year.
Figure 1. Diagrammatic representation of the myosin II protein and its subfragments. Myosin II or conventional myosin is a hexameric protein consisting of two heavy chains and two light chains. Myosin II can be proteolytically cleaved into heavy meromyosin (HMM, Cat.# MH01) and light meromyosin (LMM) by α-chymotrypsin. Heavy meromyosin consists of the myosin head subfragment-1 domain (S1), its associated light chains (essential light chains and regulatory light chains), and the coiled-coil subfragment -2 domain. Light meromyosin consists of coiledcoil protein structure. The myosin S1-subfragment is produced by papain digestion of HMM.
Protein purity is determined by scanning densitometry of Coomassie Blue stained protein on a 4-20% gradient polyacrylamide gel. Myosin protein is 95% pure (see Figure 2).
Figure 2. Myosin II protein purity determination. A 20 µg sample of MY02 was separated by electrophoresis in a 4-20% SDS-PAGE system, and stained with Coomassie Blue. Arrow indicates the myosin heavy chain (approx. 200 kDa), arrowheads indicate the RLC (approx. 20 kDa) and two ELC isoforms (approx. 25 and 17 kDa). Protein quantitation was performed using the Precision Red Protein Assay Reagent (Cat.# ADV02).
The biological activity of rabbit myosin II is determined from its rate of F-actin activated ATP hydrolysis. A standard biological assay for monitoring ATP hydrolysis by myosin consists of an in vitro F-actin ATPase assay (Cat. # BK054). Stringent quality control ensures that in the presence of F-actin, rabbit myosin will have a minimum hydrolysis rate 10 fold greater than in the absence of F-actin, which is comparable to published results.
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|Al Azzam, Omayma et al.||Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers||JoVE (Journal of Visualized Experiments)||2022||ISSN 1940--087X|
|Bashirzadeh, Yashar et al.||Encapsulated actomyosin patterns drive cell-like membrane shape changes||iScience||2022|
|Sheung, Janet Y. et al.||Motor-driven advection competes with crowding to drive spatiotemporally heterogeneous transport in cytoskeleton composites||Frontiers in Physics||2022|
|Giampazolias, Evangelos et al.||Secreted gelsolin inhibits DNGR-1-dependent cross-presentation and cancer immunity||Cell||2021||ISSN 1097-4172|
|Lee, Gloria et al.||Myosin-driven actin-microtubule networks exhibit self-organized contractile dynamics||Science Advances||2021||ISSN 2375-2548|
|Bashirzadeh, Yashar et al.||Encapsulated Actomyosin Patterns Drive Cell-Like Membrane Shape Changes||SSRN Electronic Journal||2021||Article Link|
|Lee, Gloria et al.||Active cytoskeletal composites display emergent tunable contractility and restructuring||Soft Matter||2021||ISSN 1744-6848|
|Radnai, Laszlo et al.||A semi-high-throughput adaptation of the nadh-coupled atpase assay for screening small molecule inhibitors||Journal of Visualized Experiments||2019||ISSN 1940-087X|
|Cervero, Pasquale et al.||Lymphocyte-specific protein 1 regulates mechanosensory oscillation of podosomes and actin isoform-based actomyosin symmetry breaking||Nature Communications||2018||ISSN 2041-1723|
|Ehrlicher, A. J. et al.||Mechanical strain in actin networks regulates FilGAP and integrin binding to filamin A||Nature 2011 478:7368||2011||ISSN 1476--4687|
|Rao, Mala V. et al.||The myosin Va head domain binds to the neurofilament-L rod and modulates endoplasmic reticulum (ER) content and distribution within axons||PLoS ONE||2011||ISSN 1932-6203|
|Harris, Elizabeth S. et al.||The Mouse Formin, FRLα, Slows Actin Filament Barbed End Elongation, Competes with Capping Protein, Accelerates Polymerization from Monomers, and Severs Filaments||Journal of Biological Chemistry||2004||ISSN 0021-9258|
|Gallo, Gianluca et al.||Actin turnover is required to prevent axon retraction driven by endogenous actomyosin contractility||2002||PMID 12356866|
Question 1: Does this myosin contain both light and heavy chains and all other subunits?
Answer 1: Yes, this protein is full length myosin motor protein isolated from rabbit skeletal muscle. The myosin protein contains the two heavy chains and two light chains, the essential light chain (ELC) and regulatory light chain (RLC).
Question 2: Does this myosin have ATPase activity that is activated by actin filaments?
Answer 2: Yes, the biological activity test for the myosin protein is in vitro measurement of myosin’s ATPase activity in the activating presence of actin filaments. Stringent quality control ensures that in the presence of F-actin, rabbit skeletal muscle myosin will have a minimum hydrolysis rate 10 fold greater than in the absence of F-actin.
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