title: The N- and C-Terminal Domains Differentially Contribute to the Structure and Function of Dystrophin and Utrophin Tandem Calponin-Homology Domains.
abstract: Dystrophin and utrophin are two muscle proteins involved in Duchenne/Becker muscular dystrophy. Both proteins use tandem calponin-homology (CH) domains to bind to F-actin. We probed the role of N-terminal CH1 and C-terminal CH2 domains in the structure and function of dystrophin tandem CH domain and compared with our earlier results on utrophin to understand the unifying principles of how tandem CH domains work. Actin cosedimentation assays indicate that the isolated CH2 domain of dystrophin weakly binds to F-actin compared to the full-length tandem CH domain. In contrast, the isolated CH1 domain binds to F-actin with an affinity similar to that of the full-length tandem CH domain. Thus, the obvious question is why the dystrophin tandem CH domain requires CH2, when its actin binding is determined primarily by CH1. To answer, we probed the structural stabilities of CH domains. The isolated CH1 domain is very unstable and is prone to serious aggregation. The isolated CH2 domain is very stable, similar to the full-length tandem CH domain. These results indicate that the main role of CH2 is to stabilize the tandem CH domain structure. These conclusions from dystrophin agree with our earlier results on utrophin, indicating that this phenomenon of differential contribution of CH domains to the structure and function of tandem CH domains may be quite general. The N-terminal CH1 domains primarily determine the actin binding function whereas the C-terminal CH2 domains primarily determine the structural stability of tandem CH domains, and the extent of stabilization depends on the strength of inter-CH domain interactions.
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, and ‡Program in Structural Biology and Biochemistry, University of Colorado Anschutz Medical Campus , Aurora, Colorado 80045, United States.
authors.forename: Surinder M
authors.initials: SM
authors.lastname: Singh
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, and ‡Program in Structural Biology and Biochemistry, University of Colorado Anschutz Medical Campus , Aurora, Colorado 80045, United States.
authors.forename: Swati
authors.initials: S
authors.lastname: Bandi
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, and ‡Program in Structural Biology and Biochemistry, University of Colorado Anschutz Medical Campus , Aurora, Colorado 80045, United States.
authors.forename: Krishna M G
authors.initials: KM
authors.lastname: Mallela
mesh_terms: D000199:Actins / Q000378:metabolism; D000595:Amino Acid Sequence; D055592:Biophysical Phenomena; D002135:Calcium-Binding Proteins / Q000737:chemistry*; D016189:Dystrophin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008840:Microfilament Proteins / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D040681:Structural Homology, Protein; D013816:Thermodynamics; D049411:Utrophin / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000096985:Calponins
chemical_list: D000199:Actins; D002135:Calcium-Binding Proteins; D016189:Dystrophin; D008840:Microfilament Proteins; D049411:Utrophin; D000096985:Calponins; C484258:DMD protein, human; C483599:UTRN protein, human