HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 277, Issue 40, 37272-37279, October 4, 2002

This Article Full Text Full Text (PDF) All Versions of this Article: 277/40/37272    most recent M204257200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Litowski, J. R. Articles by Hodges, R. S. Search for Related Content PubMed PubMed Citation Articles by Litowski, J. R. Articles by Hodges, R. S. Social Bookmarking                      What's this?

Designing Heterodimeric Two-stranded -Helical Coiled-coils
EFFECTS OF HYDROPHOBICITY AND -HELICAL PROPENSITY ON PROTEIN FOLDING, STABILITY, AND SPECIFICITY^*

Jennifer R. Litowski^§¶ and Robert S. Hodges^¶

From the  Department of Biochemistry, University of Alberta, Edmonton, Alberta T6G 2H7 Canada and the ^¶ Department of Biochemistry and Molecular Genetics, University of Colorado Health Sciences Center, Denver, Colorado 80262

The E/K coil, a heterodimeric coiled-coil, has been designed as a universal peptide capture and delivery system for use in applications such as biosensors and as an expression and affinity purification tag. In this design, heterodimer formation is specified through the placement of charged residues at the e and g positions of the heptad repeat such that the E coil contains all glutamic acid residues at these positions, and the K coil contains all lysine residues at these positions. The affinity and stability of the E/K coil have been modified to allow a greater range of conditions for association and dissociation. Increasing the hydrophobicity of the coiled-coil core, by substituting isoleucine for valine, gave increases in stability of 2.81 and 3.73 kcal/mol (0.47 kcal/mol/substitution). Increasing the -helical propensity of residues outside the core, by substituting alanine for serine, yielded increases in stability of 2.68 and 3.28 kcal/mol (0.41 and 0.45 kcal/mol/substitution). These sequence changes yielded a series of heterodimeric coiled-coils whose stabilities varied from 6.8 to 11.2 kcal/mol, greatly expanding their scope for use in protein engineering and biomedical applications.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				P. Q. Nguyen, S. Liu, J. C. Thompson, and J. J. Silberg Thermostability promotes the cooperative function of split adenylate kinases Protein Eng. Des. Sel., May 1, 2008; 21(5): 303 - 310. [Abstract] [Full Text] [PDF]

				J. M. Mason, M. A. Schmitz, K. M. Muller, and K. M. Arndt Semirational design of Jun-Fos coiled coils with increased affinity: Universal implications for leucine zipper prediction and design PNAS, June 13, 2006; 103(24): 8989 - 8994. [Abstract] [Full Text] [PDF]

				V. Rishi, J. Gal, D. Krylov, J. Fridriksson, M. S. Boysen, S. Mandrup, and C. Vinson SREBP-1 Dimerization Specificity Maps to Both the Helix-Loop-Helix and Leucine Zipper Domains: USE OF A DOMINANT NEGATIVE J. Biol. Chem., March 19, 2004; 279(12): 11863 - 11874. [Abstract] [Full Text] [PDF]

				D. L. Lee, C. T. Mant, and R. S. Hodges A Novel Method to Measure Self-association of Small Amphipathic Molecules: TEMPERATURE PROFILING IN REVERSED-PHASE CHROMATOGRAPHY J. Biol. Chem., June 13, 2003; 278(25): 22918 - 22927. [Abstract] [Full Text] [PDF]