{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T15:25:01Z","timestamp":1788362701174,"version":"build-2803163510"},"reference-count":48,"publisher":"American Chemical Society (ACS)","issue":"22","license":[{"start":{"date-parts":[[2015,10,14]],"date-time":"2015-10-14T00:00:00Z","timestamp":1444780800000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R01-GM068120"],"award-info":[{"award-number":["R01-GM068120"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2014,11,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Binding kinetics of the multivalent proteins peanut agglutinin (PnA) and cholera toxin B subunit (CTB) to a GM1-doped 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) lipid bilayer were investigated by both second-harmonic correlation spectroscopy (SHCS) and a traditional equilibrium binding isotherm. Adsorption and desorption rates, as well as binding affinity and binding free energy, for three bulk protein concentrations were determined by SHCS. For PnA binding to GM1, the measured adsorption rate decreased with increasing bulk PnA concentration from (3.7 \u00b1 0.3) \u00d7 106 M\u20131\u00b7s\u20131 at 0.43 \u03bcM PnA to (1.1 \u00b1 0.1) \u00d7 105 M\u20131\u00b7s\u20131 at 12 \u03bcM PnA. CTB\u2013GM1 exhibited a similar trend, decreasing from (1.0 \u00b1 0.1) \u00d7 109 M\u20131\u00b7s\u20131 at 0.5 nM CTB to (3.5 \u00b1 0.2) \u00d7 106 M\u20131\u00b7s\u20131 at 240 nM CTB. The measured desorption rates in both studies did not exhibit any dependence on initial protein concentration. As such, 0.43 \u03bcM PnA and 0.5 nM CTB had the strongest measured binding affinities, (3.7 \u00b1 0.8) \u00d7 109 M\u20131 and (2.8 \u00b1 0.5) \u00d7 1013 M\u20131, respectively. Analysis of the binding isotherm data suggests there is electrostatic repulsion between protein molecules when PnA binds GM1, while CTB\u2013GM1 demonstrates positive ligand\u2013ligand cooperativity. This study provides additional insight into the complex interactions between multivalent proteins and their ligands and showcases SHCS for examining these complex yet technologically important protein\u2013ligand complexes used in biosensors, immunoassays, and other biomedical diagnostics.<\/jats:p>","DOI":"10.1021\/ac500094v","type":"journal-article","created":{"date-parts":[[2014,10,14]],"date-time":"2014-10-14T17:29:08Z","timestamp":1413307748000},"page":"11045-11054","source":"Crossref","is-referenced-by-count":11,"title":["Determination of Multivalent Protein\u2013Ligand\nBinding Kinetics by Second-Harmonic Correlation Spectroscopy"],"prefix":"10.1021","volume":"86","author":[{"given":"Krystal L.","family":"Sly","sequence":"first","affiliation":[{"name":"University of Utah , , 315 South 1400 East , Room 2020, , ,","place":["Salt Lake City, Utah, United States, 84112"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John C.","family":"Conboy","sequence":"additional","affiliation":[{"name":"University of Utah , , 315 South 1400 East , Room 2020, , ,","place":["Salt Lake City, Utah, United States, 84112"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2014,10,14]]},"reference":[{"key":"2026090211114169000_cit1","doi-asserted-by":"crossref","first-page":"2113","DOI":"10.1039\/b405344c","volume":"2","author":"Arosio","year":"2004","journal-title":"Org. 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