{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T09:26:09Z","timestamp":1787563569820,"version":"build-2736575974"},"reference-count":55,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2014,2,12]],"date-time":"2014-02-12T00:00:00Z","timestamp":1392163200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["chemistry-europe.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["ChemPhysChem"],"published-print":{"date-parts":[[2014,4,4]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    A variety of IR\u2010active moieties with absorptions that are distinct from those of proteins have been developed as probes of local protein environments, including carbon\u2010deuterium bonds (C\uf8ffD), cyano groups (CN), and azides (N\n                    <jats:sub>3<\/jats:sub>\n                    ); however, no systematic analysis of their utility in a protein has been published. Previously, we characterized the N\u2010terminal Src homology\u20093 domain of the murine adapter protein Crk\u2010II (nSH3) with C\uf8ffD bonds site\u2010selectively incorporated throughout, and showed that it is relatively rigid and electrostatically heterogeneous and that it thermally unfolds under equilibrium conditions via a simple two\u2010state mechanism. We now report the synthesis and characterization of eight variants of nSH3 with CN and\/or N\n                    <jats:sub>3<\/jats:sub>\n                    probes at five of the same positions. In agreement with previous studies, the position\u2010dependent spectra suggest that both probes are predominantly sensitive to hydration, and not to their local electrostatic environments. Importantly, both probes also tend to significantly perturb the protein if they are not incorporated at surface\u2010exposed positions. Thus, unlike C\uf8ffD labels, which are both sensitive to their environment and non\u2010perturbative, CN and N\n                    <jats:sub>3<\/jats:sub>\n                    probes should be used with caution.\n                  <\/jats:p>","DOI":"10.1002\/cphc.201400017","type":"journal-article","created":{"date-parts":[[2014,2,12]],"date-time":"2014-02-12T08:20:53Z","timestamp":1392193253000},"page":"849-853","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":42,"title":["IR Probes of Protein Microenvironments: Utility and Potential for Perturbation"],"prefix":"10.1002","volume":"15","author":[{"given":"Ramkrishna","family":"Adhikary","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00f6rg","family":"Zimmermann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Philip 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