Unusual properties of the fungal conventional kinesin neck domain from Neurospora crassa
| Autor: | Edgar Meyhofer, Luis Moroder, Robert A. Cross, Stefan Lakämper, Günther Woehlke, Ulrike Majdic, Manfred Schliwa, Athina Kallipolitou, Dominga Deluca |
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| Rok vydání: | 2001 |
| Předmět: |
Time Factors
Protein Conformation ATPase Kinesin 13 Kinesins Biology Microtubules Models Biological Article Mass Spectrometry General Biochemistry Genetics and Molecular Biology Neurospora crassa Motor protein Adenosine Triphosphate Protein structure Cell Movement Microtubule Catalytic Domain Animals Cysteine Cloning Molecular Molecular Biology Adenosine Triphosphatases General Immunology and Microbiology Circular Dichroism General Neuroscience Temperature Processivity biology.organism_classification Protein Structure Tertiary Adenosine Diphosphate Kinetics Biochemistry Mutagenesis Site-Directed Biophysics biology.protein Kinesin Drosophila Peptides |
| Zdroj: | The EMBO Journal. 20:6226-6235 |
| ISSN: | 1460-2075 |
| DOI: | 10.1093/emboj/20.22.6226 |
| Popis: | Fungal conventional kinesins are unusually fast microtubule motor proteins. To compare the functional organization of fungal and animal conventional kinesins, a set of C-terminal deletion mutants of the Neurospora crassa conventional kinesin, NcKin, was investigated for its biochemical and biophysical properties. While the shortest, monomeric construct comprising the catalytic core and the neck-linker (NcKin343) displays very high steady-state ATPase (k(cat) = 260/s), constructs including both the full neck and adjacent hinge domains (NcKin400, NcKin433 and NcKin480) show wild-type behaviour: they are dimeric, show fast gliding and slower ATP turnover rates (k(cat) = 60-84/s), and are chemically processive. Unexpectedly, a construct (NcKin378, corresponding to Drosophila KHC381) that includes just the entire coiled-coil neck is a monomer. Its ATPase activity is slow (k(cat) = 27/s), and chemical processivity is abolished. Together with a structural analysis of synthetic neck peptides, our data demonstrate that the NcKin neck domain behaves differently from that of animal conventional kinesins and may be tuned to drive fast, processive motility. |
| Databáze: | OpenAIRE |
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