Structure and Dynamics of LC8 Complexes with KXTQT-Motif Peptides: Swallow and Dynein Intermediate Chain Compete for a Common Site

Benison, Gregory; Karplus, P. Andrew; Barbar, Elisar

doi:10.1016/j.jmb.2007.05.046

Cited by 79 publications

(117 citation statements)

References 46 publications

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“…It is possible that the lysine residues from the opposite subunit next to the phosphate group are affected by the negative charge of the phosphate group. In contrast to the structural comparison between LC8⅐ neuronal nitric oxid synthase and LC8⅐Swa complexes (42) and despite the aforementioned LC8 subunit shift, we have not observed an enlargement of the peptidebinding cleft in the Nek9 P-peptide complex. This movement in the homodimer quaternary structure upon Ser 944 phosphorylation could be significant to perturb the binding and stability of the Nek9 peptide to LC8.…”

Section: Circular Dichroism Experiments On Lc8 Binding To the Nek9contrasting

confidence: 99%

Structural Analysis of the Regulation of the DYNLL/LC8 Binding to Nek9 by Phosphorylation

Gallego

Velázquez‐Campoy

Regué

et al. 2013

Journal of Biological Chemistry

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Background: Phosphorylation regulates binding of the mitotic kinase Nek9 to LC8, controlling signal transduction through the Nek9/Nek6/7 module. Results: Crystal structures reveal the basis for the reduced interaction of LC8 for Nek9 upon phosphorylation on Nek9(Ser 944 ). Conclusion:The reduced binding affinity of Nek9 for LC8 is due to diminished enthalpic interactions. Significance: Disclosing phosphorylation as a novel mechanism that directly regulates LC8 protein-protein interactions.

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Section: Circular Dichroism Experiments On Lc8 Binding To the Nek9contrasting

confidence: 99%

Structural Analysis of the Regulation of the DYNLL/LC8 Binding to Nek9 by Phosphorylation

Gallego

Velázquez‐Campoy

Regué

et al. 2013

Journal of Biological Chemistry

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“…Previous work revealed that LC8 binds to DIC 29,30 , but the binding partner of Arl3 is unknown. To examine whether Arl3 binds either dynein, dynactin or LC8, we performed glutathione S-transferase (GST) pull-down assays using Arl3-conjugated beads.…”

Section: Resultsmentioning

confidence: 99%

Arl3 and LC8 regulate dissociation of dynactin from dynein

et al. 2014

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Cytoplasmic dynein acts as a motor for the intracellular retrograde motility of vesicles and organelles along microtubules. However, the regulatory mechanism underlying release of dynactin bound cargoes from dynein motor remains largely unknown. Here we report that ADP-ribosylation factor-like 3 (Arl3) and dynein light chain LC8 induce dissociation of dynactin from dynein. Immunoprecipitation and microtubule pull-down assays revealed that Arl3(Q71L) and LC8 facilitated detachment of dynactin from dynein. We also demonstrated Arl3(Q71L) or LC8-mediated dynactin release from a dynein-dynactin complex through trace experiments using quantum dot (Qdot)-conjugated proteins. Furthermore, we disclosed interactions of Arl3 and LC8 with dynactin and dynein, respectively, by live-cell imaging. Finally, knockdown (KD) of Arl3 and LC8 by siRNA induced abnormal localizations of dynein, dynactin and related organelles. Our findings uncovered the surprising functional relevance of GTP-bound Arl3 and LC8 for the unloading regulation of dynactin-bound cargo from dynein motor.

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“…More intensively studied DYNLL-binding proteins include neuronal nitric-oxide synthase (nNOS) (5), myoVa (2), Bcl-2-modifying factor (Bmf) (6), Bcl-2 interacting mediator (Bim) (7), dynein intermediate chain (DIC) (8), the Drosophila swallow mRNA localizing protein (9), and p21-activated protein kinase 1 (Pak1) (10,11). Several solution and crystal structures of apo-DYNLL and complexes with binding peptides have been determined (12)(13)(14)(15)(16)(17)(18)(19)(20). DYNLL has a homodimeric structure, and the bound partner peptides lie in two identical grooves formed at the dimerization interface (12)(13)(14)(15)(16)(17)(18)(19)(20).…”

Section: Lc8 Dynein Light Chain (Dynll)mentioning

confidence: 99%

“…Several solution and crystal structures of apo-DYNLL and complexes with binding peptides have been determined (12)(13)(14)(15)(16)(17)(18)(19)(20). DYNLL has a homodimeric structure, and the bound partner peptides lie in two identical grooves formed at the dimerization interface (12)(13)(14)(15)(16)(17)(18)(19)(20). Formerly, it was widely assumed that DYNLL could function as a cargo adapter on dynein and myoVa motors (6,21,22).…”

Section: Lc8 Dynein Light Chain (Dynll)mentioning

confidence: 99%

Affinity, Avidity, and Kinetics of Target Sequence Binding to LC8 Dynein Light Chain Isoforms

Radnai

Rapali

Hódi

et al. 2010

Journal of Biological Chemistry

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LC8 dynein light chain (DYNLL) is a highly conserved eukaryotic hub protein with dozens of binding partners and various functions beyond being a subunit of dynein and myosin Va motor proteins. Here, we compared the kinetic and thermodynamic parameters of binding of both mammalian isoforms, DYNLL1 and DYNLL2, to two putative consensus binding motifs (KXTQTX and XG(I/V)QVD) and report only subtle differences. Peptides containing either of the above motifs bind to DYNLL2 with micromolar affinity, whereas a myosin Va peptide (lacking the conserved Gln) and the noncanonical Pak1 peptide bind with K d values of 9 and 40 M, respectively. Binding of the KXTQTX motif is enthalpy-driven, although that of all other peptides is both enthalpy-and entropy-driven. Moreover, the KXTQTX motif shows strikingly slower off-rate constant than the other motifs. As most DYNLL partners are homodimeric, we also assessed the binding of bivalent ligands to DYNLL2. Compared with monovalent ligands, a significant avidity effect was found as follows: K d values of 37 and 3.5 nM for a dimeric myosin Va fragment and a Leu zipper dimerized KXTQTX motif, respectively. Ligand binding kinetics of DYNLL can best be described by a conformational selection model consisting of a slow isomerization and a rapid binding step. We also studied the binding of the phosphomimetic S88E mutant of DYNLL2 to the dimeric myosin Va fragment, and we found a significantly lower apparent K d value (3 M). We conclude that the thermodynamic and kinetic fine-tuning of binding of various ligands to DYNLL could have physiological relevance in its interaction network.

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Structure and Dynamics of LC8 Complexes with KXTQT-Motif Peptides: Swallow and Dynein Intermediate Chain Compete for a Common Site

Cited by 79 publications

References 46 publications

Structural Analysis of the Regulation of the DYNLL/LC8 Binding to Nek9 by Phosphorylation

Structural Analysis of the Regulation of the DYNLL/LC8 Binding to Nek9 by Phosphorylation

Arl3 and LC8 regulate dissociation of dynactin from dynein

Affinity, Avidity, and Kinetics of Target Sequence Binding to LC8 Dynein Light Chain Isoforms

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