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TitleStructure of spastin bound to a glutamate-rich peptide implies a hand-over-hand mechanism of substrate translocation.
Journal, issue, pagesJ Biol Chem, Vol. 295, Issue 2, Page 435-443, Year 2020
Publish dateJan 10, 2020
AuthorsHan Han / Heidi L Schubert / John McCullough / Nicole Monroe / Michael D Purdy / Mark Yeager / Wesley I Sundquist / Christopher P Hill /
PubMed AbstractMany members of the AAA+ ATPase family function as hexamers that unfold their protein substrates. These AAA unfoldases include spastin, which plays a critical role in the architecture of eukaryotic ...Many members of the AAA+ ATPase family function as hexamers that unfold their protein substrates. These AAA unfoldases include spastin, which plays a critical role in the architecture of eukaryotic cells by driving the remodeling and severing of microtubules, which are cytoskeletal polymers of tubulin subunits. Here, we demonstrate that a human spastin binds weakly to unmodified peptides from the C-terminal segment of human tubulin α1A/B. A peptide comprising alternating glutamate and tyrosine residues binds more tightly, which is consistent with the known importance of glutamylation for spastin microtubule severing activity. A cryo-EM structure of the spastin-peptide complex at 4.2 Å resolution revealed an asymmetric hexamer in which five spastin subunits adopt a helical, spiral staircase configuration that binds the peptide within the central pore, whereas the sixth subunit of the hexamer is displaced from the peptide/substrate, as if transitioning from one end of the helix to the other. This configuration differs from a recently published structure of spastin from , which forms a six-subunit spiral without a transitioning subunit. Our structure resembles other recently reported AAA unfoldases, including the meiotic clade relative Vps4, and supports a model in which spastin utilizes a hand-over-hand mechanism of tubulin translocation and microtubule remodeling.
External linksJ Biol Chem / PubMed:31767681 / PubMed Central
MethodsEM (single particle)
Resolution4.2 - 4.3 Å
Structure data

EMDB-20327: Spastin Hexamer in complex with substrate peptide
PDB-6pek: Structure of Spastin Hexamer (Subunit A-E) in complex with substrate peptide
PDB-6pen: Structure of Spastin Hexamer (whole model) in complex with substrate peptide
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-20805:
Spastin Hexamer (unsharpened map)
Method: EM (single particle) / Resolution: 4.3 Å

Chemicals

ChemComp-ADP:
ADENOSINE-5'-DIPHOSPHATE / ADP, energy-carrying molecule*YM

ChemComp-BEF:
BERYLLIUM TRIFLUORIDE ION

ChemComp-MG:
Unknown entry

Source
  • homo sapiens (human)
  • synthetic construct (others)
KeywordsMOTOR PROTEIN / AAA+ ATPase / Microtubule Severing

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