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Cell Rep. 2018 May 29;23(9):2795-2804. doi: 10.1016/j.celrep.2018.04.096.

Allosteric Coupling of CARMIL and V-1 Binding to Capping Protein Revealed by Hydrogen-Deuterium Exchange.

Author information

1
Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA.
2
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA.
3
Department of Chemistry, Box 1134, Washington University, One Brookings Drive, St. Louis, MO 63130, USA.
4
Department of Pathology and Immunology, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA. Electronic address: gamarasinghe@wustl.edu.
5
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine in St. Louis, St. Louis, MO 63110, USA. Electronic address: jcooper11@gmail.com.

Abstract

Actin assembly is important for cell motility. The ability of actin subunits to join or leave filaments via the barbed end is critical to actin dynamics. Capping protein (CP) binds to barbed ends to prevent subunit gain and loss and is regulated by proteins that include V-1 and CARMIL. V-1 inhibits CP by sterically blocking one binding site for actin. CARMILs bind at a distal site and decrease the affinity of CP for actin, suggested to be caused by conformational changes. We used hydrogen-deuterium exchange with mass spectrometry (HDX-MS) to probe changes in structural dynamics induced by V-1 and CARMIL binding to CP. V-1 and CARMIL induce changes in both proteins' binding sites on the surface of CP, along with a set of internal residues. Both also affect the conformation of CP's ββ subunit "tentacle," a second distal actin-binding site. Concerted regulation of actin assembly by CP occurs through allosteric couplings between CP modulator and actin binding sites.

KEYWORDS:

CARMIL; actin regulation; allostery; hydrogen-deuterium exchange; mass spectrometry

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