Structural Analysis of Human Cofilin 2/Filamentous Actin Assemblies: Atomic-Resolution Insights from Magic Angle Spinning NMR Spectroscopy

• Published in: Scientific reports 2017-03, Vol.7 (1), p.44506-44506, Article 44506
• Main Authors: Yehl, Jenna, Kudryashova, Elena, Reisler, Emil, Kudryashov, Dmitri, Polenova, Tatyana
• Format: Article
• Language: English
• Subjects: 631/535/878/1264; 631/80/128/1276; 82/6; Actin; Actin Cytoskeleton - chemistry; Actin Cytoskeleton - genetics; Actins - chemistry; Adenosine Triphosphate - chemistry; Binding Sites; Cell division; Cofilin; Cofilin 2 - chemistry; Cofilin 2 - genetics; Endocytosis; Filaments; Humanities and Social Sciences; Humans; Hydrolysis; Magnetic resonance spectroscopy; Microfilament Proteins - chemistry; Microfilament Proteins - genetics; Models, Molecular; multidisciplinary; NMR; Nuclear magnetic resonance; Nuclear Magnetic Resonance, Biomolecular; Protein Conformation; Science; Spectroscopy; Structural analysis
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep44506

Cellular actin dynamics is an essential element of numerous cellular processes, such as cell motility, cell division and endocytosis. Actin’s involvement in these processes is mediated by many actin-binding proteins, among which the cofilin family plays unique and essential role in accelerating actin treadmilling in filamentous actin (F-actin) in a nucleotide-state dependent manner. Cofilin preferentially interacts with older filaments by recognizing time-dependent changes in F-actin structure associated with the hydrolysis of ATP and release of inorganic phosphate (P i ) from the nucleotide cleft of actin. The structure of cofilin on F-actin and the details of the intermolecular interface remain poorly understood at atomic resolution. Here we report atomic-level characterization by magic angle spinning (MAS) NMR of the muscle isoform of human cofilin 2 (CFL2) bound to F-actin. We demonstrate that resonance assignments for the majority of atoms are readily accomplished and we derive the intermolecular interface between CFL2 and F-actin. The MAS NMR approach reported here establishes the foundation for atomic-resolution characterization of a broad range of actin-associated proteins bound to F-actin.