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Structural Determination of a SCAN Domain Homodimer and Heterodimer
Author(s) -
Sander Tara L.,
Peterson Francis C.,
Jensen Davin,
Heisner Alicia K.,
Hayes Paulette,
Waltner Jeanette K.,
Volkman Brian F.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a95-c
Subject(s) - dimer , crystallography , heteronuclear single quantum coherence spectroscopy , chemical shift , chemistry , two dimensional nuclear magnetic resonance spectroscopy , dihedral angle , heteronuclear molecule , monomer , nuclear magnetic resonance spectroscopy , stereochemistry , molecule , hydrogen bond , organic chemistry , polymer
The SCAN dimerization motif is found in more than 60 C 2 H 2 zinc finger proteins. The mechanism of SCAN dimerization and the full complement of protein partners are unknown. The purpose of this study was to determine the structure of a SCAN homodimer and heterodimer to identify residues at the dimer interface regulating selective assembly of SCAN proteins. The SCAN domain of the tumor suppressor gene, MZF1, self‐associates and the hypoalphalipoproteinemia susceptibility gene, ZNF202, interacts with the ZNF24 SCAN domain. Structures of the MZF1‐SCAN homodimer and ZNF202‐ZNF24 SCAN heterodimer were determined by NMR spectroscopy. Predictions of secondary structure and backbone ψ and ϕ torsion angle values were generated from a conformational database chemical shift homology search. All 1 H, 15 N and 13 C shifts were assigned using standard triple‐resonance NMR methods. The SCAN monomer is comprised of 5 helices. Inclusion of NOEs derived from heteronuclear filtered/edited NOESY spectra confirmed that MZF1‐SCAN is a domain‐swapped homodimer with a binding interface in agreement with other published studies. The structure revealed that helix 3 (H3) is at the center of the dimer interface. Residues connecting H1 and H2 form extensive contacts across the dimer to H3, H5, and the other monomer. GST pulldowns of recombinant SCAN proteins confirmed structural predictions and identified novel protein partners. These studies provide valuable insight into the mechanism of SCAN dimerization and protein partner selection for this large family of transcription factors.