{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T07:15:03Z","timestamp":1788333303749,"version":"build-2803163510"},"reference-count":80,"publisher":"American Chemical Society (ACS)","issue":"43","license":[{"start":{"date-parts":[[2015,10,7]],"date-time":"2015-10-07T00:00:00Z","timestamp":1444176000000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000001","name":"National Science Foundation","doi-asserted-by":"publisher","award":["OCI-1053575"],"award-info":[{"award-number":["OCI-1053575"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["P30 EY00331"],"award-info":[{"award-number":["P30 EY00331"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000053","name":"National Eye Institute","doi-asserted-by":"publisher","award":["R01 EY05216"],"award-info":[{"award-number":["R01 EY05216"]}],"id":[{"id":"10.13039\/100000053","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["GM075962"],"award-info":[{"award-number":["GM075962"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,10,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Site-directed spin labeling in combination with EPR is a powerful method for providing distances on the nm scale in biological systems. The most popular strategy, double electron\u2013electron resonance (DEER), is carried out at cryogenic temperatures (50\u201380 K) to increase the short spin\u2013spin relaxation time (T2) upon which the technique relies. A challenge is to measure long-range distances (20\u201360 \u00c5) in proteins near physiological temperatures. Toward this goal we are investigating an alternative approach based on the distance-dependent enhancement of spin\u2013lattice relaxation rate (T1\u20131) of a nitroxide spin label by a paramagnetic metal. With a commonly used nitroxide side chain (R1) and Cu2+, it has been found that interspin distances \u226425 \u00c5 can be determined in this way (Jun et al. Biochemistry\u00a02006, 45, 11666). Here, the upper limit of the accessible distance is extended to \u224840 \u00c5 using spin labels with long T1, a high-affinity 5-residue Cu2+ binding loop inserted into the protein sequence, and pulsed saturation recovery to measure relaxation enhancement. Time-domain Cu2+ electron paramagnetic resonance, quantum mechanical calculations, and molecular dynamics simulations provide information on the structure and geometry of the Cu2+ loop and indicate that the metal ion is well-localized in the protein. An important aspect of these studies is that both Cu2+\/nitroxide DEER at cryogenic temperatures and T1 relaxation measurements at room temperature can be carried out on the same sample, allowing both validation of the relaxation method and assessment of the effect of freezing on protein structure.<\/jats:p>","DOI":"10.1021\/ja5083206","type":"journal-article","created":{"date-parts":[[2014,10,7]],"date-time":"2014-10-07T09:56:52Z","timestamp":1412675812000},"page":"15356-15365","source":"Crossref","is-referenced-by-count":47,"title":["Long-Range\nDistance Measurements in Proteins at Physiological\nTemperatures Using Saturation Recovery EPR Spectroscopy"],"prefix":"10.1021","volume":"136","author":[{"given":"Zhongyu","family":"Yang","sequence":"first","affiliation":[{"name":"University of California , , , ,","place":["Los Angeles, California, United States, 90095"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gonzalo","family":"Jime\u0301nez-Ose\u0301s","sequence":"additional","affiliation":[{"name":"University of California , , , ,","place":["Los Angeles, California, United States, 90095"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos J.","family":"Lo\u0301pez","sequence":"additional","affiliation":[{"name":"University of California , , , ,","place":["Los Angeles, California, United States, 90095"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael D.","family":"Bridges","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K. 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