title: Resonance Assignments and Secondary Structure Analysis of Dynein Light Chain 8 by Magic Angle Spinning NMR Spectroscopy.
abstract: Dynein light chain LC8 is the smallest subunit of the dynein motor complex and has been shown to play important roles in both dynein dependent and dynein independent physiological functions via its interaction with a number of its binding partners. It has also been linked to pathogenesis including roles in viral infections and tumorigenesis. Structural information for LC8-target proteins is critical to understanding the underlying function of LC8 in these complexes. However, some LC8-target interactions are not amenable for structural characterization by conventional structural biology techniques due to their large size, low solubility and crystallization difficulties. Here, we report magic angle spinning (MAS) NMR studies of the homodimeric apo-LC8 protein as a first effort in addressing more complex, multi-partner LC8-based protein assemblies. We have established site-specific backbone and side chain resonance assignments for the majority of the residues of LC8, and show TALOS+ predicted torsion angles ϕ and ψ in close agreement with most residues in the published LC8 crystal structure. Data obtained through these studies will provide the first step toward using MAS NMR to examine the LC8 structure, which will eventually be used to investigate protein-protein interactions in larger systems, which cannot be determined by conventional structural studies.
authors.affiliation: Department of Chemistry and Biochemistry, University of Delaware, Newark, DE 19716, United States.
authors.forename: Shangjin
authors.initials: S
authors.lastname: Sun
authors.forename: Andrew H
authors.initials: AH
authors.lastname: Butterworth
authors.forename: Sivakumar
authors.initials: S
authors.lastname: Paramasivam
authors.forename: Si
authors.initials: S
authors.lastname: Yan
authors.forename: Christine M
authors.initials: CM
authors.lastname: Lightcap
authors.forename: John C
authors.initials: JC
authors.lastname: Williams
authors.forename: Tatyana
authors.initials: T
authors.lastname: Polenova