Journal of the American Chemical Society
Article
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monitoring tool revealed melting temperatures of Tm > 80 °C
for 4SR and Tm = 58 °C for 4RR (Figure 4c). These relative
stabilities of both cross-linked peptides are significantly higher
than those found for the self-assembled homotrimeric helices
formed by all of the previously examined CMPs. As further
references we also determined the melting temperatures of the
triple helices formed by CMPs 5, 6R, 6S, 7R, and 7S and found
Tm values in the range 31−39 °C (Table S1). Thus, the
covalent oxime cross-links led to an increase of the relative
stabilities of the triple helices by 20 to >40 °C. The difference
in melting temperatures of ΔTm > 20 °C between CMPs 4SR
and 4RR shows that the absolute stereochemistry of the linkage
point significantly influences the thermal stability of the cross-
linked triple helix. As predicted, the cross-linked triple helix
with (4S)-configured Aop residues in the Xaa positions is
significantly more stable compared to the isomeric trimer with a
(4R)Aop residue. Thus, the unfavorable sterics of the
substituent in the homotrimeric triple helix formed by CMPs
2YR and 2XS are overcome by their integration into a covalent
tether.
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CONCLUSIONS
■
In summary, we have developed an oxime linker that covalently
connects short collagen peptides between Aop and Alp proline
residues placed in coplanar Xaa and Yaa positions within triple
helices. This linkage enabled access to nondistorted triple
helices with remarkable thermal stabilities. The study put forth
the value of Aop as a handle for cross-linking and for the facile
functionalization of collagen peptides. The work also high-
lighted the utility of this residue for the adjustment of triple-
helical stabilities by the choice of the stereochemistry at C(4).
Together, these findings open new opportunities for the design
of functional collagen-based materials.
(8) Erdmann, R. S.; Wennemers, H. J. Am. Chem. Soc. 2010, 132,
13957.
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Ed. 2014, 53, 10340.
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50, 6835. (b) Erdmann, R. S.; Wennemers, H. J. Am. Chem. Soc. 2012,
134, 17117.
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Chem. Soc. 2002, 124, 14085. (b) Horng, J.-C.; Hawk, A. J.; Zhao, Q.;
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(c) Byrne, C.; McEwan, P. A.; Emsley, J.; Fischer, P. M.; Chan, W. C.
Chem. Commun. 2011, 47, 2589. (d) Melacini, G.; Feng, Y.; Goodman,
M. J. Am. Chem. Soc. 1996, 118, 10359.
ASSOCIATED CONTENT
* Supporting Information
The Supporting Information is available free of charge on the
■
S
(13) (a) Ottl, J.; Moroder, L. J. Am. Chem. Soc. 1999, 121, 653.
(b) Kotch, F. W.; Raines, R. T. Proc. Natl. Acad. Sci. U. S. A. 2006, 103,
Details on the syntheses and analyses of the presented
compounds as well as on the simulations (PDF)
3028. (c) Bouleg
Moroder, L. Antioxid. Redox Signaling 2008, 10, 113. (d) Boudko, S.
̀
ue, C.; Musiol, H.-J.; Gotz, M. G.; Renner, C.;
̈
AUTHOR INFORMATION
Corresponding Author
ORCID
P.; Engel, J.; Okuyama, K.; Mizuno, K.; Bachinger, H. P.; Schumacher,
M. A. J. Biol. Chem. 2008, 283, 32580. (e) Tanrikulu, I. C.; Raines, R.
T. J. Am. Chem. Soc. 2014, 136, 13490.
(14) Persikov, A. V.; Ramshaw, J. A.; Kirkpatrick, A.; Brodsky, B.
Biochemistry 2000, 39, 14960.
̈
■
(15) (a) Bella, B.; Eaton, M.; Brodsky, B.; Berman, H. M. Science
Notes
1994, 266, 75. (b) Okuyama, K.; Miyama, K.; Mizuno, K.; Bachinger,
̈
H. P. Biopolymers 2012, 97, 607.
(16) For original work on Aop, see: Liu, F.; Stephen, A. G.; Fisher, R.
J.; Burke, T. R. Bioorg. Med. Chem. Lett. 2008, 18, 1096. See also:
Pandey, A. K.; Naduthambi, D.; Thomas, K. M.; Zondlo, N. J. J. Am.
Chem. Soc. 2013, 135, 4333.
(17) Dirksen, A.; Hackeng, T. M.; Dawson, P. E. Angew. Chem., Int.
Ed. 2006, 45, 7581.
The authors declare no competing financial interest.
ACKNOWLEDGMENTS
■
We thank the Fonds National de la Recherche Luxembourg
(FNR) for an AFR Ph.D. Fellowship for N.H. and the Swiss
National Science Foundation (grants SNF 200021_159858
(H.W.) and 200021_159713 (S.R.)) for financial support.
(18) For reviews, see: (a) Ulrich, S.; Boturyn, D.; Marra, A.;
Renaudet, O.; Dumy, P. Chem. - Eur. J. 2014, 20, 34. (b) Collins, J.;
Xiao, Z.; Mullner, M.; Connal, L. A. Polym. Chem. 2016, 7, 3812.
̈
(c) Agten, S. M.; Dawson, P. E.; Hackeng, T. M. J. Pept. Sci. 2016, 22,
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