Crystallization, X-ray characterization and selenomethionine phasing of Mlc1p bound to IQ motifs from myosin V

Acta Crystallogr D Biol Crystallogr. 2002 Oct;58(Pt 10 Pt 2):1882-5. doi: 10.1107/s0907444902013951. Epub 2002 Sep 28.

Abstract

Mlc1p is a calmodulin-like protein from the budding yeast Saccharomyces cerevisiae, where it has been identified as a subunit of a class V myosin, Myo2p, and a binding partner of an IQGAP-like protein, Iqg1p. Through its interactions with these two proteins, Mlc1p plays a role in polarized growth and cytokinesis. Mlc1p has been crystallized in complexes with four different IQ target motifs from the neck region of Myo2p: IQ2, IQ3, IQ4 and IQ2-IQ3 (referred to as IQ2,3). Electron-density maps for two of the complexes (Mlc1p-IQ4 and Mlc1p-IQ2,3) were obtained from multiple anomalous dispersion (MAD) experiments based on selenomethionine derivatives. The other two structures (Mlc1p-IQ2 and Mlc1p-IQ3) were determined by molecular replacement using the partially refined structure of Mlc1p-IQ2,3 as a search model.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Binding Sites
  • Cell Division
  • Cloning, Molecular
  • Escherichia coli
  • Models, Molecular
  • Molecular Sequence Data
  • Myosin Light Chains / chemistry*
  • Myosin Light Chains / isolation & purification
  • Myosin Light Chains / metabolism
  • Myosin Type V / chemistry*
  • Myosin Type V / metabolism
  • Protein Conformation
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / isolation & purification
  • Saccharomyces cerevisiae Proteins / metabolism
  • X-Ray Diffraction / methods

Substances

  • MLC1 protein, S cerevisiae
  • Myosin Light Chains
  • Recombinant Proteins
  • Saccharomyces cerevisiae Proteins
  • Myosin Type V

Associated data

  • SWISSPROT/P53141