Oligomerization properties of GCN4 leucine zipper e and g position mutants

Xiangang Zeng, Hai Zhu, Hilal A. Lashuel, James C. Hu

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Putative intersubunit electrostatic interactions between charged amine acids on the surfaces of the dimer interfaces of leucine zippers (g-e' ion pairs) have been implicated as determinants of dimerization specificity. To evaluate the importance of these ionic interactions in determining the specificity of dimer formation, we constructed a pool of >65,000 GCN4 leucine zipper mutants in which all the e and g positions are occupied by different combinations of alanine, glutamic acid, lysine, or threonine. The oligomerization properties of these mutants were evaluated based on the phenotypes of cells expressing λ repressor-leucine zipper fusion proteins. About 90% of the mutants do not form stable homooligomers. Surprisingly, approximately 8% of the mutant sequences have phenotypes consistent with the formation of higher-order (>dimer) oligomers, which can be classified into three types based on sequence features. The oligomerization states of mutants from two of these types were determined by characterizing purified fusion proteins. The Type I mutant behaved as a tetramer under all tested conditions, whereas the Type III mutant formed a variety of higher-order oligomers, depending on the solution conditions. Stable homodimers comprise less than 3% of the pool; several g-e' positions in these mutants could form attractive ion pairs. Putative repulsive ion pairs are not found among the homodimeric mutants. However, patterns of charged residues at the e and g positions do not seem to be sufficient to predict either homodimer or heterodimer formation among the mutants.

Original languageEnglish
Pages (from-to)2218-2226
Number of pages9
JournalProtein Science
Volume6
Issue number10
Publication statusPublished - 1 Oct 1997
Externally publishedYes

Fingerprint

Leucine Zippers
Oligomerization
Dimers
Ions
Oligomers
Fusion reactions
Phenotype
Dimerization
Threonine
Coulomb interactions
Static Electricity
Alanine
Lysine
Amines
Glutamic Acid
Proteins
Cells
Acids

Keywords

  • Dimerization specificity
  • Leucine zippers
  • Protein structure
  • Recombinant fusion proteins
  • Site-directed mutagenesis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Zeng, X., Zhu, H., Lashuel, H. A., & Hu, J. C. (1997). Oligomerization properties of GCN4 leucine zipper e and g position mutants. Protein Science, 6(10), 2218-2226.

Oligomerization properties of GCN4 leucine zipper e and g position mutants. / Zeng, Xiangang; Zhu, Hai; Lashuel, Hilal A.; Hu, James C.

In: Protein Science, Vol. 6, No. 10, 01.10.1997, p. 2218-2226.

Research output: Contribution to journalArticle

Zeng, X, Zhu, H, Lashuel, HA & Hu, JC 1997, 'Oligomerization properties of GCN4 leucine zipper e and g position mutants', Protein Science, vol. 6, no. 10, pp. 2218-2226.
Zeng X, Zhu H, Lashuel HA, Hu JC. Oligomerization properties of GCN4 leucine zipper e and g position mutants. Protein Science. 1997 Oct 1;6(10):2218-2226.
Zeng, Xiangang ; Zhu, Hai ; Lashuel, Hilal A. ; Hu, James C. / Oligomerization properties of GCN4 leucine zipper e and g position mutants. In: Protein Science. 1997 ; Vol. 6, No. 10. pp. 2218-2226.
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