Nuclear export inhibition through covalent conjugation and hydrolysis of Leptomycin B by CRM1

Proc Natl Acad Sci U S A. 2013 Jan 22;110(4):1303-8. doi: 10.1073/pnas.1217203110. Epub 2013 Jan 7.

Abstract

The polyketide natural product Leptomycin B inhibits nuclear export mediated by the karyopherin protein chromosomal region maintenance 1 (CRM1). Here, we present 1.8- to 2.0-Å-resolution crystal structures of CRM1 bound to Leptomycin B and related inhibitors Anguinomycin A and Ratjadone A. Structural and complementary chemical analyses reveal an unexpected mechanism of inhibition involving covalent conjugation and CRM1-mediated hydrolysis of the natural products' lactone rings. Furthermore, mutagenesis reveals the mechanism of hydrolysis by CRM1. The nuclear export signal (NES)-binding groove of CRM1 is able to drive a chemical reaction in addition to binding protein cargoes for transport through the nuclear pore complex.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acrylates / chemistry
  • Acrylates / pharmacology
  • Active Transport, Cell Nucleus / drug effects*
  • Amino Acid Substitution
  • Crystallography, X-Ray
  • Fatty Acids, Unsaturated / chemistry
  • Fatty Acids, Unsaturated / metabolism
  • Fatty Acids, Unsaturated / pharmacology
  • Humans
  • Hydrolysis
  • Karyopherins / antagonists & inhibitors
  • Karyopherins / chemistry
  • Karyopherins / genetics
  • Karyopherins / metabolism*
  • Models, Anatomic
  • Mutagenesis, Site-Directed
  • Nuclear Export Signals / genetics
  • Protein Conformation
  • Receptors, Cytoplasmic and Nuclear / antagonists & inhibitors
  • Receptors, Cytoplasmic and Nuclear / chemistry
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Static Electricity
  • Triazoles / chemistry
  • Triazoles / pharmacology

Substances

  • Acrylates
  • Fatty Acids, Unsaturated
  • KPT-185
  • Karyopherins
  • Nuclear Export Signals
  • Receptors, Cytoplasmic and Nuclear
  • Saccharomyces cerevisiae Proteins
  • Triazoles
  • exportin 1 protein
  • leptomycin B

Associated data

  • PDB/4HAT
  • PDB/4HAU
  • PDB/4HAV
  • PDB/4HAW
  • PDB/4HAX
  • PDB/4HAY
  • PDB/4HAZ
  • PDB/4HB0
  • PDB/4HB2
  • PDB/4HB3
  • PDB/4HB4