The Dictyostelium class I myosin, MyoD, contains a novel light chain that lacks high-affinity calcium-binding sites.
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MlcD is a 16 kDa calmodulin-like protein that co-purifies with MyoD as two copies per heavy chain.
"MyoD, a long-tailed class I myosin, co-purified with two copies of a 16 kDa light chain"
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MlcD has four EF-hands but only retains low-affinity calcium binding due to degenerate calcium-coordinating residues in EF-hands 2-4.
"MlcD comprises four EF-hands; however, EF-hands 2-4 contain mutations in key Ca2+-co-ordinating residues that would be predicted to impair Ca2+ binding"
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MlcD binds calcium with a Kd of 52 uM, far above physiological calcium concentrations.
"yielding apparent dissociation constants ( K'(d)) of 52 microM for Ca2+ and 450 microM for Mg2+"
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MlcD cannot function as a physiological calcium sensor.
"The low affinity of MlcD for Ca2+ indicates that it cannot function as a sensor of physiological Ca2+"
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MlcD binding to MyoD is calcium-insensitive.
"Ca2+ did not affect the binding of MlcD to MyoD or to either of the two MyoD IQ (Ile-Gln) motifs"
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MlcD specifically associates with MyoD and not with MyoB or MyoC.
"FLAG-MlcD expressed in Dictyostelium formed a complex with MyoD, but not with the two other long-tailed Dictyostelium myosin I isoenzymes, MyoB and MyoC"