Structural basis for capping protein sequestration by myotrophin (V-1).
Capping protein (CP) is a ubiquitously expressed, heterodimeric 62-kDa protein that binds the barbed end of the actin filament with high affinity to block further filament elongation. Myotrophin (V-1) is a 13-kDa ankyrin repeat-containing protein that binds CP tightly, sequestering it in a totally inactive complex in vitro. Here, we elucidate the molecular interaction between CP and V-1 by NMR. Specifically, chemical shift mapping and intermolecular paramagnetic relaxation enhancement experiments reveal that the ankyrin loops of V-1, which are essential for V-1/CP interaction, bind the basic patch near the joint of the alpha tentacle of CP shown previously to drive most of the association of CP with and affinity for the barbed end. Consistently, site-directed mutagenesis of CP shows that V-1 and the strong electrostatic binding site for CP on the barbed end compete for this basic patch on CP. These results can explain how V-1 inactivates barbed end capping by CP and why V-1 is incapable of uncapping CP-capped actin filaments, the two signature biochemical activities of V-1.
Zwolak
Adam
A
Laboratory of Molecular Biophysics, HLBI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Fujiwara
Ikuko
I
Hammer
John A
JA
Tjandra
Nico
N
D051344:Actin Capping Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002645:Chickens; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D011485:Protein Binding / Q000235:genetics / Q000502:physiology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011817:Rabbits
D051344:Actin Capping Proteins; D036341:Intercellular Signaling Peptides and Proteins; C065700:myotrophin