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TitleStructure of γ-tubulin small complex based on a cryo-EM map, chemical cross-links, and a remotely related structure.
Journal, issue, pagesJ Struct Biol, Vol. 194, Issue 3, Page 303-310, Year 2016
Publish dateMar 8, 2016
AuthorsCharles H Greenberg / Justin Kollman / Alex Zelter / Richard Johnson / Michael J MacCoss / Trisha N Davis / David A Agard / Andrej Sali /
PubMed AbstractModeling protein complex structures based on distantly related homologues can be challenging due to poor sequence and structure conservation. Therefore, utilizing even low-resolution experimental ...Modeling protein complex structures based on distantly related homologues can be challenging due to poor sequence and structure conservation. Therefore, utilizing even low-resolution experimental data can significantly increase model precision and accuracy. Here, we present models of the two key functional states of the yeast γ-tubulin small complex (γTuSC): one for the low-activity "open" state and another for the higher-activity "closed" state. Both models were computed based on remotely related template structures and cryo-EM density maps at 6.9Å and 8.0Å resolution, respectively. For each state, extensive sampling of alignments and conformations was guided by the fit to the corresponding cryo-EM density map. The resulting good-scoring models formed a tightly clustered ensemble of conformations in most regions. We found significant structural differences between the two states, primarily in the γ-tubulin subunit regions where the microtubule binds. We also report a set of chemical cross-links that were found to be consistent with equilibrium between the open and closed states. The protocols developed here have been incorporated into our open-source Integrative Modeling Platform (IMP) software package (http://integrativemodeling.org), and can therefore be applied to many other systems.
External linksJ Struct Biol / PubMed:26968363 / PubMed Central
MethodsEM (helical sym.)
Resolution6.9 - 8 Å
Structure data

PDB-5flz:
Cryo-EM structure of gamma-TuSC oligomers in a closed conformation
Method: ELECTRON MICROSCOPY / Resolution: 6.9 Å

PDB-5fm1:
Structure of gamma-tubulin small complex based on a cryo-EM map, chemical cross-links, and a remotely related structure
Method: ELECTRON MICROSCOPY / Resolution: 8.0 Å

Source
  • saccharomyces cerevisiae (brewer's yeast)
KeywordsCELL CYCLE / MICROTUBULE NUCLEATION / MICROTUBULE / NUCLEATION / TUBULIN / FILAMENT

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