In vitro reconstitution of a transport complex containing Rab27a, melanophilin and myosin Va.
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.
Wu
Xufeng
X
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.
Sakamoto
Take
T
Zhang
Fang
F
Sellers
James R
JR
Hammer
John A
JA
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001693:Biological Transport, Active; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D020460:Melanosomes / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D046912:Multiprotein Complexes; D018995:Myosin Heavy Chains / Q000235:genetics / Q000378:metabolism*; D024701:Myosin Type V / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000075745:rab27 GTP-Binding Proteins
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C435724:Mlph protein, mouse; D046912:Multiprotein Complexes; C513082:Myo5a protein, mouse; D011993:Recombinant Fusion Proteins; D000075745:rab27 GTP-Binding Proteins; D049452:Green Fluorescent Proteins; D024701:Myosin Type V; C512962:Rab27a protein, mouse; D018995:Myosin Heavy Chains; D020691:rab GTP-Binding Proteins