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While prior studies identified slow conformational exchange at the interdomain interface, these minor states could not be structurally linked to activation because their resonances broaden beyond detection upon receptor binding. Here, we use multinuclear NMR spectroscopy to characterize the intrinsic conformational landscape of full\u2010length, human arrestin\u20102 in solution. We identify two distinct, pre\u2010existing conformational equilibria that mirror key steps of the receptor\u2010driven activation process. First, a slow exchange process populates a receptor\u2010bound\u2010like, interdomain\u2010twisted conformation at physiological temperatures. In parallel, a faster, globally distributed equilibrium populates a state consistent with C\u2010terminal tail release. Dynamic analyses reveal localized rigidification in the receptor\u2010bound\u2010like minor states despite arrestin's overall flexibility, while backbone relaxation data indicate widespread \u03bcs\u2013ms conformational exchange. Together, these results demonstrate that arrestin\u20102 acts as a preorganized scaffold that intrinsically samples receptor\u2010binding relevant conformations in the absence of binding partners. This provides a solution\u2010state framework for arrestin pre\u2010activation and establishes a dynamic fingerprint for future ligand\u2010dependent studies.<\/jats:p>","DOI":"10.1002\/pro.70627","type":"journal-article","created":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T13:16:52Z","timestamp":1778851012000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Intrinsic conformational equilibria position arrestin\u20102 for activation"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-5121-8881","authenticated-orcid":false,"given":"Tucker J.","family":"Shriver","sequence":"first","affiliation":[{"name":"Department of Pharmacology Northwestern University Feinberg School of Medicine  Chicago Illinois USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-8684-0832","authenticated-orcid":false,"given":"Kerem","family":"Kahraman","sequence":"additional","affiliation":[{"name":"Department of Pharmacology Northwestern University Feinberg School of Medicine  Chicago Illinois USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5954-3240","authenticated-orcid":false,"given":"Mingzhe","family":"Pan","sequence":"additional","affiliation":[{"name":"Department of Cellular and Integrative Physiology University of Texas Health San Antonio  San Antonio Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1595-431X","authenticated-orcid":false,"given":"\u00c7a\u011fda\u015f","family":"Da\u011f","sequence":"additional","affiliation":[{"name":"Nanofabrication and Nanocharacterization Center for Scientific and Technological Advance Research (n<sup>2<\/sup>STAR) Ko\u00e7 University  Istanbul Turkiye"},{"name":"Ko\u00e7 University Isbank Center for Infectious Diseases (KUISCID) Ko\u00e7 University  Istanbul T\u00fcrkiye"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7700-5745","authenticated-orcid":false,"given":"Marco","family":"Tonelli","sequence":"additional","affiliation":[{"name":"National Magnetics Resonance Facility at Madison, Department of Biochemistry University of Wisconsin\u2010Madison  Madison Wisconsin USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0529-0399","authenticated-orcid":false,"given":"Scott A.","family":"Robson","sequence":"additional","affiliation":[{"name":"Department of Pharmacology Northwestern University Feinberg School of Medicine  Chicago Illinois USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3740-9999","authenticated-orcid":false,"given":"Joshua J.","family":"Ziarek","sequence":"additional","affiliation":[{"name":"Department of Pharmacology Northwestern University Feinberg School of Medicine  Chicago Illinois 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