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The paramagnetic effects manifested in NMR spectroscopy generally contain paramagnetic relaxation enhancement, pseudocontact shifts (PCSs), and residual dipolar coupling (RDC), and these effects correlate closely with the flexibility of protein\u2013tag conjugates. The rigidity of the paramagnetic tag is greatly important in decoding the structural details of macromolecular complexes, because paramagnetic averaging reduces the PCSs and RDCs. Here we show that the dynamic exchange of the metal chelating moiety is a key factor in determining the rigidity of the paramagnetic tag in the protein conjugates. Decreasing the conformational exchange rates in the metal chelating moiety greatly minimizes the paramagnetic averaging and thus increases PCSs and RDCs. This effect has been demonstrated in an open-chain tag, Py-l-Cys-DTPA, which generates large PCSs and RDCs that are comparable to those of the reported cyclic DOTA-like tags. The proposed route offers a unique way to design suitable paramagnetic tags for applications in biological systems.<\/jats:p>","DOI":"10.1021\/acs.jpclett.0c02196","type":"journal-article","created":{"date-parts":[[2020,10,27]],"date-time":"2020-10-27T17:19:05Z","timestamp":1603819145000},"page":"9493-9500","source":"Crossref","is-referenced-by-count":11,"title":["Dynamic Exchange of the Metal Chelating Moiety: A\nKey Factor in Determining the Rigidity of Protein\u2013Tag Conjugates\nin Paramagnetic NMR"],"prefix":"10.1021","volume":"11","author":[{"given":"Jia-Liang","family":"Chen","sequence":"first","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300071"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Li","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300071"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xia-Yan","family":"Li","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300071"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3051-0047","authenticated-orcid":true,"given":"Xun-Cheng","family":"Su","sequence":"additional","affiliation":[{"name":"Nankai University , , ,","place":["Tianjin, China, 300071"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,10,27]]},"reference":[{"key":"2026081723153449300_cit1","doi-asserted-by":"publisher","first-page":"211","DOI":"10.1016\/j.pnmrs.2019.06.003","article-title":"Magnetic Susceptibility\nand Paramagnetism-based\nNMR","volume":"114\u2013115","author":"Parigi","year":"2019","journal-title":"Prog. 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Commun."},{"key":"2026081723153449300_cit29","doi-asserted-by":"publisher","first-page":"4877","DOI":"10.1021\/jm901876r","article-title":"Exofacial\nProtein Thiols as a Route for the Internalization of Gd\u00ad(III)-Based\nComplexes for Magnetic Resonance Imaging Cell Labeling","volume":"53","author":"Digilio","year":"2010","journal-title":"J. Med. Chem."},{"key":"2026081723153449300_cit30","doi-asserted-by":"publisher","first-page":"323","DOI":"10.1002\/ijch.198400055","article-title":"Heavy Metal Ionophores: Correlations Among Structural\nParameters of Complexed Nonpeptide Polyamino Acids","volume":"24","author":"Stezowski","year":"1984","journal-title":"Isr. J. Chem."},{"key":"2026081723153449300_cit31","doi-asserted-by":"publisher","first-page":"151","DOI":"10.1042\/bj2990151","article-title":"Synthetic, Structural and Biological Studies of the Ubiquitin System:\nthe Total Chemical Synthesis of Ubiquitin","volume":"299","author":"Ramage","year":"1994","journal-title":"Biochem.\nJ."},{"key":"2026081723153449300_cit32","doi-asserted-by":"publisher","first-page":"179","DOI":"10.1007\/s10858-008-9249-z","article-title":"Numbat: an\nInteractive Software Tool for Fitting \u0394\u03c7-Tensors to Molecular\nCoordinates using Pseudocontact Shifts","volume":"41","author":"Schmitz","year":"2008","journal-title":"J. Biomol.\nNMR"},{"key":"2026081723153449300_cit33","doi-asserted-by":"publisher","first-page":"3752","DOI":"10.1021\/ja4132642","article-title":"Improved Cross\nValidation of a Static Ubiquitin Structure Derived from High Precision\nResidual Dipolar Couplings Measured in a Drug-Based Liquid Crystalline\nPhase","volume":"136","author":"Maltsev","year":"2014","journal-title":"J. Am. Chem. Soc."},{"key":"2026081723153449300_cit34","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1023\/A:1011206132740","article-title":"A Novel Interactive\nTool for Rigid-Body Modeling of Multi-Domain Macromolecules using\nResidual Dipolar Couplings","volume":"20","author":"Dosset","year":"2001","journal-title":"J. Biomol. 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Acta"},{"key":"2026081723153449300_cit46","doi-asserted-by":"publisher","first-page":"5889","DOI":"10.1002\/anie.200352234","article-title":"Towards\nthe Rational\nDesign of Magnetic Resonance Imaging Contrast Agents: Isolation of\nthe two Coordination Isomers of Lanthanide DOTA-Type Complexes","volume":"42","author":"Woods","year":"2003","journal-title":"Angew. Chem., Int. Ed."},{"key":"2026081723153449300_cit47","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1002\/(SICI)1099-0682(200001)2000:1&lt;75::AID-EJIC75&gt;3.0.CO;2-#","article-title":"Solution Equilibria in YbDOTMA, a\nChiral Analogue of one of the Most Successful Contrast Agents for\nMRI, GdDOTA","volume":"1","author":"Di\nBari","year":"2000","journal-title":"Eur. J. Inorg. 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