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TitleInsights into the Distinct Mechanisms of Action of Taxane and Non-Taxane Microtubule Stabilizers from Cryo-EM Structures.
Journal, issue, pagesJ Mol Biol, Vol. 429, Issue 5, Page 633-646, Year 2017
Publish dateMar 10, 2017
AuthorsElizabeth H Kellogg / Nisreen M A Hejab / Stuart Howes / Peter Northcote / John H Miller / J Fernando Díaz / Kenneth H Downing / Eva Nogales /
PubMed AbstractA number of microtubule (MT)-stabilizing agents (MSAs) have demonstrated or predicted potential as anticancer agents, but a detailed structural basis for their mechanism of action is still lacking. ...A number of microtubule (MT)-stabilizing agents (MSAs) have demonstrated or predicted potential as anticancer agents, but a detailed structural basis for their mechanism of action is still lacking. We have obtained high-resolution (3.9-4.2Å) cryo-electron microscopy (cryo-EM) reconstructions of MTs stabilized by the taxane-site binders Taxol and zampanolide, and by peloruside, which targets a distinct, non-taxoid pocket on β-tubulin. We find that each molecule has unique distinct structural effects on the MT lattice structure. Peloruside acts primarily at lateral contacts and has an effect on the "seam" of heterologous interactions, enforcing a conformation more similar to that of homologous (i.e., non-seam) contacts by which it regularizes the MT lattice. In contrast, binding of either Taxol or zampanolide induces MT heterogeneity. In doubly bound MTs, peloruside overrides the heterogeneity induced by Taxol binding. Our structural analysis illustrates distinct mechanisms of these drugs for stabilizing the MT lattice and is of relevance to the possible use of combinations of MSAs to regulate MT activity and improve therapeutic potential.
External linksJ Mol Biol / PubMed:28104363 / PubMed Central
MethodsEM (helical sym.)
Resolution3.5 Å
Structure data

EMDB-8320, PDB-5syc:
Near-atomic resolution cryo-EM reconstruction of peloruside-stabilized microtubule
Method: EM (helical sym.) / Resolution: 3.5 Å

EMDB-8321, PDB-5sye:
Near-atomic resolution cryo-EM reconstruction of doubly bound Taxol- and peloruside-stabilized microtubule
Method: EM (helical sym.) / Resolution: 3.5 Å

EMDB-8322, PDB-5syf:
High-resolution cryo-EM reconstruction of Taxol-stabilized microtubule
Method: EM (helical sym.) / Resolution: 3.5 Å

EMDB-8323, PDB-5syg:
Cryo-EM reconstruction of zampanolide-bound microtubule
Method: EM (helical sym.) / Resolution: 3.5 Å

Chemicals

ChemComp-GTP:
GUANOSINE-5'-TRIPHOSPHATE / GTP, energy-carrying molecule*YM

ChemComp-MG:
Unknown entry

ChemComp-GDP:
GUANOSINE-5'-DIPHOSPHATE / GDP, energy-carrying molecule*YM

ChemComp-POU:
Peloruside A

ChemComp-TA1:
TAXOL / medication, chemotherapy*YM

ChemComp-ZPN:
(2Z,4E)-N-[(S)-[(1S,2E,5S,8E,10Z,17S)-3,11-dimethyl-19-methylidene-7,13-dioxo-6,21-dioxabicyclo[15.3.1]henicosa-2,8,10-trien-5-yl](hydroxy)methyl]hexa-2,4-dienamide

Source
  • sus scrofa (pig)
KeywordsSTRUCTURAL PROTEIN / peloruside / microtubule / Taxol / zampanolide

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