In vitro reconstitution of a transport complex containing Rab27a, melanophilin and myosin Va |
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| Authors: | Wu Xufeng Sakamoto Take Zhang Fang Sellers James R Hammer John A |
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| Affiliation: | Laboratory of Cell Biology, National Heart, Lung and Blood Institute, National Institutes of Health, Building 50 Room 2523 NIH 9000 Rockvill, Bethesda, MD 20892-8017, USA. |
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| Abstract: | Rab27a and melanophilin (Mlph) are required in vivo to form a melanosome receptor for myosin Va in which Rab27a anchored in the melanosome membrane recruits Mlph, which in turn recruits myosin Va. Here, we show by reconstitution using purified proteins that Rab27a and Mlph are sufficient to form a transport complex with myosin Va in vitro. These results suggest that additional proteins are not required in vivo for assembly of the myosin Va receptor, although other proteins may associate with this tripartite complex to regulate its activity and/or to assist Rab27a in anchoring the complex to the melanosome membrane. |
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| Keywords: | ABS, actin binding site DA, dominant active GFP, green fluorescent protein melanophilin, Mlph TIRFM, total internal reflection fluorescence microscopy WT, wild-type |
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