Surface salt bridges stabilize the GCN4 leucine zipper

Erik J. Spek, Au H. Bui, Min Lu, Neville R. Kallenbach

Research output: Contribution to journalArticle

Abstract

We present a study of the role of salt bridges in stabilizing a simplified tertiary structural motif, the coiled-coil. Changes in GCN4 sequence have been engineered that introduce trial patterns of single and multiple salt bridges at solvent exposed sites. At the same sites, a set of alanine mutants was generated to provide a reference for thermodynamic analysis of the salt bridges. Introduction of three alanines stabilizes the dimer by 1.1 kcal/mol relative to the wild-type. An arrangement corresponding to a complex type of salt bridge involving three groups stabilizes the dimer by 1.7 kcal/mol, an apparent elevation of the melting temperature relative to wild type of about 22 °C. While identifying local from nonlocal contributions to protein stability is difficult, stabilizing interactions can be identified by use of cycles. Introduction of alanines for side chains of lower helix propensity and complex salt bridges both stabilize the coiled- coil, so that combining the two should yield melting temperatures substantially higher than the starting species, approaching those of thermophilic sequences.

Original languageEnglish (US)
Pages (from-to)2431-2437
Number of pages7
JournalProtein Science
Volume7
Issue number11
StatePublished - Nov 1998

Fingerprint

Leucine Zippers
Salts
Alanine
Dimers
Freezing
Melting point
Temperature
Protein Stability
Thermodynamics
Proteins

Keywords

  • GCN4
  • Leucine zipper
  • Salt bridge
  • Thermal stability

ASJC Scopus subject areas

  • Biochemistry

Cite this

Spek, E. J., Bui, A. H., Lu, M., & Kallenbach, N. R. (1998). Surface salt bridges stabilize the GCN4 leucine zipper. Protein Science, 7(11), 2431-2437.

Surface salt bridges stabilize the GCN4 leucine zipper. / Spek, Erik J.; Bui, Au H.; Lu, Min; Kallenbach, Neville R.

In: Protein Science, Vol. 7, No. 11, 11.1998, p. 2431-2437.

Research output: Contribution to journalArticle

Spek, EJ, Bui, AH, Lu, M & Kallenbach, NR 1998, 'Surface salt bridges stabilize the GCN4 leucine zipper', Protein Science, vol. 7, no. 11, pp. 2431-2437.
Spek EJ, Bui AH, Lu M, Kallenbach NR. Surface salt bridges stabilize the GCN4 leucine zipper. Protein Science. 1998 Nov;7(11):2431-2437.
Spek, Erik J. ; Bui, Au H. ; Lu, Min ; Kallenbach, Neville R. / Surface salt bridges stabilize the GCN4 leucine zipper. In: Protein Science. 1998 ; Vol. 7, No. 11. pp. 2431-2437.
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