Journal article
An engineered leucine zipper a position mutant with an unusual three-state unfolding pathway
Protein science, Vol.10(1), pp.24-33
01/01/2001
DOI: 10.1110/ps.30901
PMCID: PMC2249852
PMID: 11266591
Abstract
The leucine zipper is a dimeric coiled-coil structural motif consisting of four to six heptad repeats, designated (
abcdefg
)
n
. In the GCN4 leucine zipper,
a
position 16 in the third heptad is occupied by an Asn residue whereas the other
a
positions are Val residues. Recently, we have constructed variants of the GCN4 leucine zipper in which the
a
position Val residues were replaced by Ile. The folding and unfolding of the wild-type GCN4 leucine zipper and the Val to Ile variant both adhere to a simple two-state mechanism. In this study, another variant of the GCN4 leucine zipper was constructed by moving the single Asn residue from
a
position 16 to
a
position 9. This switch causes the thermal unfolding of the GCN4 leucine zipper to become three state. The unfolding pathway of this variant was determined by thermal denaturation, limited proteinase K digestion, and sedimentation equilibrium analysis. Our data are consistent with a model in which the variant first unfolds from its N terminus and changes the oligomerization specificity from a native dimer to a partially unfolded intermediate containing a mixture of dimers and trimers and then completely unfolds to unstructured monomers.
Details
- Title: Subtitle
- An engineered leucine zipper a position mutant with an unusual three-state unfolding pathway
- Creators
- Hai Zhu - Texas A&M UniversityScott A. Celinski - Texas A&M UniversityJ. Martin Scholtz - Texas A&M UniversityJames C. Hu - Texas A&M University
- Resource Type
- Journal article
- Publication Details
- Protein science, Vol.10(1), pp.24-33
- DOI
- 10.1110/ps.30901
- PMID
- 11266591
- PMCID
- PMC2249852
- NLM abbreviation
- Protein Sci
- ISSN
- 0961-8368
- eISSN
- 1469-896X
- Publisher
- Cold Spring Harbor Laboratory Press
- Language
- English
- Date published
- 01/01/2001
- Academic Unit
- Research Administration; Pharmaceutical Sciences and Experimental Therapeutics; Biochemistry and Molecular Biology; Chemistry
- Record Identifier
- 9984288721702771
Metrics
26 Record Views