The Complex Interplay between the Neck and Hinge Domains in Kinesin-1 Dimerization and Motor Activity
| Autor: | Manfred Schliwa, Lucia Driller, Friederike Bathe, Guenther Woehlke, Renate Dombi, Katrin Hahlen |
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| Rok vydání: | 2005 |
| Předmět: |
Proline
Molecular Sequence Data Mutant Hinge Kinesins Sequence alignment macromolecular substances Biology Conserved sequence Neurospora crassa Structure-Activity Relationship Humans Structure–activity relationship Amino Acid Sequence Disulfides Protein Structure Quaternary Molecular Biology Peptide sequence Conserved Sequence Tryptophan Articles Cell Biology biology.organism_classification Protein Structure Tertiary Kinetics Biochemistry Mutation Biophysics Kinesin Dimerization Sequence Alignment |
| Zdroj: | Molecular Biology of the Cell. 16:3529-3537 |
| ISSN: | 1939-4586 1059-1524 |
| DOI: | 10.1091/mbc.e04-11-0957 |
| Popis: | Kinesin-1 dimerizes via the coiled-coil neck domain. In contrast to animal kinesins, neck dimerization of the fungal kinesin-1 NcKin requires additional residues from the hinge. Using chimeric constructs containing or lacking fungal-specific elements, the proximal part of the hinge was shown to stabilize the neck coiled-coil conformation in a complex manner. The conserved fungal kinesin hinge residue W384 caused neck coiled-coil formation in a chimeric NcKin construct, including parts of the human kinesin-1 stalk. The stabilizing effect was retained in a NcKinW384F mutant, suggesting important π -stacking interactions. Without the stalk, W384 was not sufficient to induce coiled-coil formation, indicating that W384 is part of a cluster of several residues required for neck coiled-coil folding. A W384-less chimera of NcKin and human kinesin possessed a non–coiled-coil neck conformation and showed inhibited activity that could be reactivated when artificial interstrand disulfide bonds were used to stabilize the neck coiled-coil conformation. On the basis of yeast two-hybrid data, we propose that the proximal hinge can bind kinesin's cargo-free tail domain and causes inactivation of kinesin by disrupting the neck coiled-coil conformation. |
| Databáze: | OpenAIRE |
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