Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2.
| dc.contributor.author | Milam, Sara L | |
| dc.contributor.author | Erickson, Harold P | |
| dc.date.accessioned | 2018-04-01T14:48:11Z | |
| dc.date.available | 2018-04-01T14:48:11Z | |
| dc.date.issued | 2013-08 | |
| dc.date.updated | 2018-04-01T14:48:10Z | |
| dc.description.abstract | FtsZ from most bacteria assembles rapidly in vitro, reaching a steady-state plateau in 5-10 s after addition of GTP. A recent study used a novel dynamic light-scattering technique to assay the assembly of FtsZ from Caulobacter crescentus (CcFtsZ) and reported that assembly required 10 min, ∼100 times slower than for related bacteria. Previous studies had indicated normal, rapid assembly of CcFtsZ. We have reinvestigated the assembly kinetics using a mutant L72W, where assembly of subunits into protofilaments results in a significant increase in tryptophan fluorescence. We found that assembly reached a plateau in 5-10 s and showed no change in the following 10 min. This was confirmed by 90° light scattering and negative-stain electron microscopy. The very slow kinetics in the dynamic light-scattering study may be related to a refractory state induced when the FtsZ protein is stored without nucleotide, a phenomenon that we had observed in a previous study of EcFtsZ. We conclude that CcFtsZ is not an outlier, but shows rapid assembly kinetics similar to FtsZ from related bacteria. | |
| dc.identifier.issn | 1083-351X | |
| dc.identifier.issn | 1083-351X | |
| dc.identifier.uri | ||
| dc.language | eng | |
| dc.publisher | American Society for Biochemistry & Molecular Biology (ASBMB) | |
| dc.relation.ispartof | The Journal of biological chemistry | |
| dc.relation.isversionof | 10.1074/jbc.m113.491845 | |
| dc.subject | Caulobacter crescentus | |
| dc.subject | Magnesium | |
| dc.subject | GTP Phosphohydrolases | |
| dc.subject | Bacterial Proteins | |
| dc.subject | Cytoskeletal Proteins | |
| dc.subject | Guanosine Triphosphate | |
| dc.subject | Negative Staining | |
| dc.subject | Hydrolysis | |
| dc.subject | Kinetics | |
| dc.subject | Hydrogen-Ion Concentration | |
| dc.subject | Fluorescence | |
| dc.subject | Scattering, Radiation | |
| dc.title | Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2. | |
| dc.type | Journal article | |
| pubs.issue | 33 | |
| pubs.organisational-group | School of Medicine | |
| pubs.organisational-group | Duke | |
| pubs.organisational-group | Duke Cancer Institute | |
| pubs.organisational-group | Institutes and Centers | |
| pubs.organisational-group | Biochemistry | |
| pubs.organisational-group | Basic Science Departments | |
| pubs.organisational-group | Cell Biology | |
| pubs.publication-status | Published | |
| pubs.volume | 288 |