Purification and Structural Analysis of SUN and KASH Domain Proteins

Methods Enzymol. 2016;569:63-78. doi: 10.1016/bs.mie.2015.08.011. Epub 2015 Sep 6.

Abstract

Molecular tethers span the nuclear envelope to mechanically connect the cytoskeleton and nucleoskeleton. These bridge-like tethers, termed linkers of nucleoskeleton and cytoskeleton (LINC) complexes, consist of SUN proteins at the inner nuclear membrane and KASH proteins at the outer nuclear membrane. LINC complexes are central to a variety of cell activities including nuclear positioning and mechanotransduction, and LINC-related abnormalities are associated with a spectrum of tissue-specific diseases, termed laminopathies or envelopathies. Protocols used to study the biochemical and structural characteristics of core elements of SUN-KASH complexes are described here to facilitate further studies in this new field of cell biology.

Keywords: KASH peptide; LINC complex; Nesprin; Nuclear envelope; SUN domain; SUN protein.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Sequence
  • Cell Cycle Proteins / chemistry
  • Cell Cycle Proteins / isolation & purification*
  • Chromatography, Affinity
  • Chromatography, Gel
  • Crystallography, X-Ray
  • Escherichia coli
  • Humans
  • Intracellular Signaling Peptides and Proteins / chemistry
  • Intracellular Signaling Peptides and Proteins / isolation & purification*
  • Membrane Proteins / chemistry
  • Membrane Proteins / isolation & purification*
  • Molecular Sequence Data
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / isolation & purification*
  • Peptide Fragments / chemistry
  • Peptide Fragments / isolation & purification
  • Protein Structure, Tertiary

Substances

  • Cell Cycle Proteins
  • Intracellular Signaling Peptides and Proteins
  • KASH5 protein, human
  • Membrane Proteins
  • Nuclear Proteins
  • Peptide Fragments
  • SUN2 protein, human