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Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2.

Publication ,  Journal Article
Milam, SL; Erickson, HP
Published in: J Biol Chem
August 16, 2013

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.

Duke Scholars

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

August 16, 2013

Volume

288

Issue

33

Start / End Page

23675 / 23679

Location

United States

Related Subject Headings

  • Scattering, Radiation
  • Negative Staining
  • Magnesium
  • Kinetics
  • Hydrolysis
  • Hydrogen-Ion Concentration
  • Guanosine Triphosphate
  • GTP Phosphohydrolases
  • Fluorescence
  • Cytoskeletal Proteins
 

Citation

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Milam, S. L., & Erickson, H. P. (2013). Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2. J Biol Chem, 288(33), 23675–23679. https://doi.org/10.1074/jbc.M113.491845
Milam, Sara L., and Harold P. Erickson. “Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2.J Biol Chem 288, no. 33 (August 16, 2013): 23675–79. https://doi.org/10.1074/jbc.M113.491845.
Milam SL, Erickson HP. Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2. J Biol Chem. 2013 Aug 16;288(33):23675–9.
Milam, Sara L., and Harold P. Erickson. “Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2.J Biol Chem, vol. 288, no. 33, Aug. 2013, pp. 23675–79. Pubmed, doi:10.1074/jbc.M113.491845.
Milam SL, Erickson HP. Rapid in vitro assembly of Caulobacter crescentus FtsZ protein at pH 6.5 and 7.2. J Biol Chem. 2013 Aug 16;288(33):23675–23679.

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

August 16, 2013

Volume

288

Issue

33

Start / End Page

23675 / 23679

Location

United States

Related Subject Headings

  • Scattering, Radiation
  • Negative Staining
  • Magnesium
  • Kinetics
  • Hydrolysis
  • Hydrogen-Ion Concentration
  • Guanosine Triphosphate
  • GTP Phosphohydrolases
  • Fluorescence
  • Cytoskeletal Proteins