Charge-pair interactions between acidic and basic residues on the surface of collagen can promote stability as well as control specificity of molecular recognition. Heterotrimeric collagen peptides have been engineered de novo using either rational or computational methods, which in both cases optimize networks of favorable charge-pair interactions in the target structure. Less understood is the role of electrostatic repulsion between groups of like charge in destabilizing structure or directing molecular recognition. To study this, we apply a “charge crowding” approach, where repulsive interactions between multiple aspartate side chains are found to destabilize the homotrimer states in triple helical peptide system and can be utilized to promote the formation of heterotrimers. Neutralizing surface charge by increasing salt concentration or decreasing pH can enhance homotrimer stability, confirming the role of charge crowding on the destabilization of homotrimers via electrostatic repulsion. Charge crowding may be used in conjunction with other approaches to create specific collagen heterotrimers.
References
[1]
Di Lullo, G.A.; Sweeney, S.M.; Korkko, J.; Ala-Kokko, L.; San Antonio, J.D. Mapping the ligand-binding sites and disease-associated mutations on the most abundant protein in the human, type I collagen. J. Biol. Chem. 2002, 277, 4223–4231.
[2]
Kadler, K.E.; Holmes, D.F.; Trotter, J.A.; Chapman, J.A. Collagen fibril formation. Biochem. J. 1996, 316, 1–11.
[3]
Ramachandran, G.N.; Kartha, G. Structure of collagen. Nature 1955, 176, 593–595, doi:10.1038/176593a0.
[4]
Rich, A.; Crick, F.H. The molecular structure of collagen. J. Mol. Biol. 1961, 3, 483–506, doi:10.1016/S0022-2836(61)80016-8.
[5]
Salem, G.; Traub, W. Conformational implications of amino acid sequence regularities in collagen. FEBS Lett. 1975, 51, 94–99, doi:10.1016/0014-5793(75)80861-1.
[6]
Ramshaw, J.A.; Shah, N.K.; Brodsky, B. Gly-X-Y tripeptide frequencies in collagen: A context for host-guest triple-helical peptides. J. Struct. Biol. 1998, 122, 86–91.
[7]
Mason, J.M.; Hagemann, U.B.; Arndt, K.M. Role of hydrophobic and electrostatic interactions in coiled coil stability and specificity. Biochemistry 2009, 48, 10380–10388.
[8]
O’Shea, E.K.; Lumb, K.J.; Kim, P.S. Peptide “Velcro”: Design of a heterodimeric coiled coil. Curr. Biol. 1993, 3, 658–667, doi:10.1016/0960-9822(93)90063-T.
[9]
Nautiyal, S.; Woolfson, D.N.; King, D.S.; Alber, T. A designed heterotrimeric coiled coil. Biochemistry 1995, 34, 11645–11651.
[10]
Kohn, W.D.; Monera, O.D.; Kay, C.M.; Hodges, R.S. The effects of interhelical electrostatic repulsions between glutamic acid residues in controlling the dimerization and stability of two-stranded alpha-helical coiled-coils. J. Biol. Chem. 1995, 270, 25495–25506.
[11]
Kohn, W.D.; Kay, C.M.; Hodges, R.S. Protein destabilization by electrostatic repulsions in the two-stranded alpha-helical coiled-coil/leucine zipper. Protein Sci. 1995, 4, 237–250.
[12]
Kohn, W.D.; Kay, C.M.; Hodges, R.S. Positional dependence of the effects of negatively charged Glu side chains on the stability of two-stranded alpha-helical coiled-coils. J. Pept. Sci. 1997, 3, 209–223, doi:10.1002/(SICI)1099-1387(199705)3:3<209::AID-PSC102>3.0.CO;2-S.
Fallas, J.A.; Dong, J.; Tao, Y.J.; Hartgerink, J.D. Structural insights into charge pair interactions in triple helical collagen-like proteins. J. Biol. Chem. 2012, 287, 8039–8047.
[16]
Gauba, V.; Hartgerink, J.D. Surprisingly high stability of collagen ABC heterotrimer: Evaluation of side chain charge pairs. J. Am. Chem. Soc. 2007, 129, 15034–15041, doi:10.1021/ja075854z.
Xu, F.; Zahid, S.; Silva, T.; Nanda, V. Computational design of a collagen A:B:C-type heterotrimer. J. Am. Chem. Soc. 2011, 133, 15260–15263, doi:10.1021/ja205597g.
[19]
Parmar, A.S.; Zahid, S.; Belure, S.V.; Young, R.; Hasan, N.; Nanda, V. Design of net-charged abc-type collagen heterotrimers. J. Struct. Biol. 2013, doi:10.1016/j.jsb.2013.04.006.
[20]
Giddu, S.; Xu, F.; Nanda, V. Sequence recombination improves target specificity in a redesigned collagen peptide abc-type heterotrimer. Proteins 2013, 81, 386–393, doi:10.1002/prot.24194.
[21]
Xu, F.; Zhang, L.; Koder, R.L.; Nanda, V. De novo self-assembling collagen heterotrimers using explicit positive and negative design. Biochemistry 2010, 49, 2307–2316, doi:10.1021/bi902077d.
[22]
Parmar, A.S.; Nunes, A.M.; Baum, J.; Brodsky, B. A peptide study of the relationship between the collagen triple-helix and amyloid. Biopolymers 2012, 97, 795–806, doi:10.1002/bip.22070.
[23]
Persikov, A.V.; Ramshaw, J.A.; Brodsky, B. Prediction of collagen stability from amino acid sequence. J. Biol. Chem. 2005, 280, 19343–19349, doi:10.1074/jbc.M501657200.
[24]
Bretscher, L.E.; Jenkins, C.L.; Taylor, K.M.; DeRider, M.L.; Raines, R.T. Conformational stability of collagen relies on a stereoelectronic effect. J. Am. Chem. Soc. 2001, 123, 777–778.
