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. 2010 Jun 9;132(22):7586-8.
doi: 10.1021/ja100080q.

Side-chain pairing preferences in the parallel coiled-coil dimer motif: insight on ion pairing between core and flanking sites

Jay D Steinkruger  1 , Derek N Woolfson, Samuel H Gellman
Affiliations

Side-chain pairing preferences in the parallel coiled-coil dimer motif: insight on ion pairing between core and flanking sites

Jay D Steinkruger et al. J Am Chem Soc. .

Abstract

A new strategy for rapid evaluation of sequence-stability relationships in the parallel coiled-coil motif is described. The experimental design relies upon thiol-thioester exchange equilibria, an approach that is particularly well suited to examination of heterodimeric systems. Our model system has been benchmarked by demonstrating that it can quantitatively reproduce previously reported trends in interhelical a-a' side-chain pairing preferences at the coiled-coil interface. This new tool has been used to explore the role of Coulombic interactions between a core position on one helix and a flanking position on the other helix (a-g'). This type of interhelical contact has received relatively little attention to date. Our results indicate that such interactions can influence coiled-coil partner preferences.

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Figures

Figure 1
Figure 1
(a) Primary sequence of CT-C' (X = Ψ =Ile). (b) Helical wheel diagram showing the proposed helical regions of CT-C'. Guest sites are highlighted in dashed boxes (c) Cartoon illustrating the thiol-thioester exchange process.
Figure 2
Figure 2
(a) Thermodynamic data (ΔGCC) values determined from thioester exchange of CT-C' mutants. Assays were conducted in 50 mM sodium phosphate buffer (pH 7) with 2 mM TCEP at 25 °C. (b) Correlation diagram comparing Δ(ΔG) data from the Vinson laboratory, and our work using thioester exchange. Δ(ΔG) values are normalized to the Ala-Ala homotypic pairing.
Figure 3
Figure 3
Partial helical net diagrams and ΔGCC values depicting for different a–g' or a'–g ion pairs.
See this image and copyright information in PMC

References

    1. Grigoryan G, Keating AE. Curr. Opin. Struct. Biol. 2008;18:477. - PMC - PubMed
    2. Woolfson DN. Adv. Protein Chem. 2005;70:79. - PubMed
    3. Mason JM, Arndt KM. ChemBioChem. 2004;5:170. - PubMed
    4. Parry DAD, Bruce Fraser RD, Squire JM. J. Struct. Biol. 2008;163:258. - PubMed
    5. Cohen C, Parry DAD. Proteins. 1990;7:1. - PubMed
    1. Crick FHS. Acta Crystallogr. 1953;6:689.
    1. Acharya A, Rishi V, Vinson C. Biochemistry. 2006;45:11324. - PubMed
    2. Acharya A, Ruvinov SB, Gal J, Moll JR, Vinson C. Biochemistry. 2002;41:14122. - PubMed
    3. Moitra J, Szilak L, Krylov D, Vinson C. Biochemistry. 1997;36:12567. - PubMed
    4. Tripet B, Wagschal K, Lavigne P, Mant CT, Hodges RS. J. Mol. Biol. 2000;300:377. - PubMed
    5. Wagschal K, Tripet B, Lavigne P, Mant C, Hodges RS. Protein Sci. 1999;8:2312. - PMC - PubMed
    6. Ryan SJ, Kennan AJ. J. Am. Chem. Soc. 2007;129:10255. - PubMed
    1. Bromley EH, Channon K, Moutevelis E, Woolfson DN. ACS Chem. Biol. 2008;3:38. - PubMed
    1. Woll MG, Gellman SH. J. Am. Chem. Soc. 2004;126:11172. - PubMed
    2. Hadley EB, Gellman SH. J. Am. Chem. Soc. 2006;128:16444. - PubMed
    3. Hadley EB, Testa OD, Woolfson DN, Gellman SH. Proc. Natl. Acad. Sci. U.S.A. 2008;105:530. - PMC - PubMed

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