title: Flexibility in the N-terminal actin-binding domain: clues from in silico mutations and molecular dynamics.
abstract: Dystrophin is a long, rod-shaped cytoskeleton protein implicated in muscular dystrophy (MDys). Utrophin is the closest autosomal homolog of dystrophin. Both proteins have N-terminal actin-binding domain (N-ABD), a central rod domain and C-terminal region. N-ABD, composed of two calponin homology (CH) subdomains joined by a helical linker, harbors a few disease causing missense mutations. Although the two proteins share considerable homology (>72%) in N-ABD, recent structural and biochemical studies have shown that there are significant differences (including stability, mode of actin-binding) and their functions are not completely interchangeable. In this investigation, we have used extensive molecular dynamics simulations to understand the differences and the similarities of these two proteins, along with another actin-binding protein, fimbrin. In silico mutations were performed to identify two key residues that might be responsible for the dynamical difference between the molecules. Simulation points to the inherent flexibility of the linker region, which adapts different conformations in the wild type dystrophin. Mutations T220V and G130D in dystrophin constrain the flexibility of the central helical region, while in the two known disease-causing mutants, K18N and L54R, the helicity of the region is compromised. Phylogenetic tree and sequence analysis revealed that dystrophin and utrophin genes have probably originated from the same ancestor. The investigation would provide insight into the functional diversity of two closely related proteins and fimbrin, and contribute to our understanding of the mechanism of MDys.
authors.affiliation: Department of Biochemistry, Bose Institute, Kolkata, West Bengal, 700054, India.
authors.forename: Devlina
authors.initials: D
authors.lastname: Chakravarty
authors.forename: Soumyananda
authors.initials: S
authors.lastname: Chakraborti
authors.forename: Pinak
authors.initials: P
authors.lastname: Chakrabarti
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D016189:Dystrophin / Q000737:chemistry / Q000145:classification / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D009136:Muscular Dystrophies; D009154:Mutation; D010802:Phylogeny; D011485:Protein Binding; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D016415:Sequence Alignment; D049411:Utrophin / Q000737:chemistry / Q000378:metabolism
chemical_list: D000199:Actins; D016189:Dystrophin; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D049411:Utrophin; C026742:plastin
keywords: actin-binding proteins; dimer vs monomer; muscular dystrophy; oligomeric state; open and closed states; structural flexibility