A Buried Polar Residue in the Hydrophobic Interface of the Coiled-coil Peptide, GCN4-p1, Plays a Thermodynamic, not a Kinetic Role in Folding

Knappenberger, Jane A.; Smith, Jennifer E.; Thorpe, Sarah H.; Zitzewitz, Jill A.; Matthews, C. Robert

doi:10.1016/s0022-2836(02)00592-2

Cited by 37 publications

(39 citation statements)

References 22 publications

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“…As seen by the appearance of a single minimum at 203 nm in the data collected at elevated temperatures, the structure of PF4 ZIP reversibly changes to random-coil at elevated temperature. Maximum mean residue ellipticity values at 222 nm (MRE 222 ) were measured to be -30,470 deg cm 2 dmol -1 for the PF4 ZIP at 15 °C, consistent with reported values ranging from -25,000 to -35,000 deg cm 2 dmol -1 for various GCN4-mimic peptides in PBS at 15 °C [42,43].…”

Section: Oligomerization Of Pf4 Zipsupporting

confidence: 86%

Manipulation of hydrogel assembly and growth factor delivery via the use of peptide–polysaccharide interactions

Zhang

Furst

Kiick

2006

Journal of Controlled Release

116

131

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The design of materials in which assembly, mechanical response, and biological properties are controlled by protein-polysaccharide interactions could provide materials that mimic the biological environment and find use in the delivery of growth factors. In the investigations reported here, a heparin-binding, coiled-coil peptide, PF4 ZIP , was employed to mediate the assembly of heparinized polymers. The heparin-binding affinity of this peptide was compared with that of other heparinbinding peptides (HBP) via heparin-sepharose chromatography and surface plasmon resonance (SPR) experiments. Results from these experiments indicate that PF4 ZIP demonstrates a higher heparin-binding affinity and heparin association rate when compared to the heparin-binding domains of antithrombin III (ATIII) and heparin-interacting protein (HIP). Viscoelastic hydrogels were formed upon the association of PF4 ZIP -functionalized star poly(ethylene glycol) (PEG-PF4 ZIP ) with low-molecular-weight heparin-functionalized star PEG (PEG-LMWH). The viscoelastic properties of the hydrogels can be altered via variations in the ratio of LMWH to PF4 ZIP . bFGF release from these gels have also been investigated. Comparison of the bFGF release profiles with the hydrogel erosion profiles indicates that bFGF delivery from this class of hydrogels is mainly an erosioncontrolled process and the rates of bFGF release can be modulated via choice of HBP or via variations in the mechanical properties of the hydrogels. Manipulation of hydrogel physical properties and erosion profiles will provide novel materials for controlled growth factor delivery and other biomedical applications.

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Section: Oligomerization Of Pf4 Zipsupporting

confidence: 86%

Manipulation of hydrogel assembly and growth factor delivery via the use of peptide–polysaccharide interactions

Zhang

Furst

Kiick

2006

Journal of Controlled Release

116

131

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“…S1). The positions of the intermediate states correspond well to the positions of asparagine residues in the sequence, which are known to destabilize the coiled coil (14,16).…”

Section: Resultsmentioning

confidence: 76%

Full distance-resolved folding energy landscape of one single protein molecule

Gebhardt

Bornschlögl

Rief

2010

Proc. Natl. Acad. Sci. U.S.A.

195

246

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Kinetic bulk and single molecule folding experiments characterize barrier properties but the shape of folding landscapes between barrier top and native state is difficult to access. Here, we directly extract the full free energy landscape of a single molecule of the GCN4 leucine zipper using dual beam optical tweezers. To this end, we use deconvolution force spectroscopy to follow an individual molecule's trajectory with high temporal and spatial resolution. We find a heterogeneous energy landscape of the GCN4 leucine zipper domain. The energy profile is divided into two stable C-terminal heptad repeats and two less stable repeats at the N-terminus. Energies and transition barrier positions were confirmed by single molecule kinetic analysis. We anticipate that deconvolution sampling is a powerful tool for the model-free investigation of protein energy landscapes.leucine zipper | force spectroscopy | optical tweezers | protein folding | deconvolution

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“…To gain a better biophysical understanding of N@d layers in coiled coils, we studied their effect in a well-understood model system, the GCN4-p1-N16V leucine zipper. In this GCN4 variant, the core asparagine in position a of the third heptad is substituted with valine, resulting in an increased stability but decreased structural specificity, as this protein forms mixtures of dimers and trimers (15). Into this background, we introduced the N@d motifs VxxNxxx, VxxNTxx, IxxNxxx, and IxxNTxx in 1 (M1), 2 (M2), or 3 (M3) consecutive heptads (Fig.…”

Section: Resultsmentioning

confidence: 99%

A coiled-coil motif that sequesters ions to the hydrophobic core

Hartmann¹,

Ridderbusch²,

Zeth³

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Most core residues of coiled coils are hydrophobic. Occasional polar residues are thought to lower stability, but impart structural specificity. The coiled coils of trimeric autotransporter adhesins (TAAs) are conspicuous for their large number of polar residues in position d of the core, which often leads to their prediction as natively unstructured regions. The most frequent residue, asparagine (N@d), can occur in runs of up to 19 consecutive heptads, frequently in the motif [I/V]xxNTxx. In the Salmonella TAA, SadA, the core asparagines form rings of interacting residues with the following threonines, grouped around a central anion. This conformation is observed generally in N@d layers from trimeric coiled coils of known structure. Attempts to impose a different register on the motif show that the asparagines orient themselves specifically into the core, even against conflicting information from flanking domains. When engineered into the GCN4 leucine zipper, N@d layers progressively destabilized the structure, but zippers with 3 N@d layers still folded at high concentration. We propose that N@d layers maintain the coiled coils of TAAs in a soluble, export-competent state during autotransport through the outer membrane. More generally, we think that polar motifs that are both periodic and conserved may often reflect special folding requirements, rather than an unstructured state of the mature proteins.ion coordination ͉ protein export ͉ trimeric autotransporter adhesin ͉ polar core residues

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A Buried Polar Residue in the Hydrophobic Interface of the Coiled-coil Peptide, GCN4-p1, Plays a Thermodynamic, not a Kinetic Role in Folding

Cited by 37 publications

References 22 publications

Manipulation of hydrogel assembly and growth factor delivery via the use of peptide–polysaccharide interactions

Manipulation of hydrogel assembly and growth factor delivery via the use of peptide–polysaccharide interactions

Full distance-resolved folding energy landscape of one single protein molecule

A coiled-coil motif that sequesters ions to the hydrophobic core

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