{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T09:35:52Z","timestamp":1790760952594,"version":"4.1.0"},"posted":{"date-parts":[[2023,8,28]]},"group-title":"Biochemistry","reference-count":41,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,8,28]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Aggregation of antibody light chain proteins is associated with the progressive disease light chain amyloidosis. Patient-derived amyloid fibrils are formed from light chain variable domain residues in non-native conformations, highlighting a requirement that light chains unfold from their native structures in order to aggregate. However, mechanistic studies of amyloid formation have primarily focused on the self-assembly of natively unstructured peptides, and the role of native state unfolding is less well understood. Using a well-studied light chain variable domain protein known as WIL, which readily aggregates\n                  <jats:italic>in vitro<\/jats:italic>\n                  under conditions where the native state predominates, we asked how the protein concentration and addition of pre-formed fibril \u201cseeds\u201d alter the kinetics of aggregation. Monitoring aggregation with thioflavin T fluorescence revealed a distinctly non-linear dependence on concentration, with a maximum aggregation rate observed at 8 \u00b5M protein. This behavior is consistent with formation of alternate aggregate structures in the early phases of amyloid formation. Addition of N- or C-terminal peptide tags, which did not greatly affect the folding or stability of the protein, altered the concentration dependence of aggregation. Aggregation rates increased in the presence of pre-formed seeds, but this effect did not eliminate the delay before aggregation and became saturated when the proportion of seeds added was greater than 1 in 1600. The complexity of aggregation observed\n                  <jats:italic>in vitro<\/jats:italic>\n                  highlights how multiple species may contribute to amyloid pathology in patients.\n                <\/jats:p>","DOI":"10.1101\/2023.08.28.555139","type":"posted-content","created":{"date-parts":[[2023,8,29]],"date-time":"2023-08-29T01:55:11Z","timestamp":1693274111000},"source":"Crossref","is-referenced-by-count":2,"title":["Kinetic evidence for multiple aggregation pathways in antibody light chain variable domains"],"prefix":"10.64898","author":[{"given":"Sherry","family":"Wong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madeline E.","family":"West","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1275-2010","authenticated-orcid":false,"given":"Gareth J.","family":"Morgan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2023090103250959000_2023.08.28.555139v1.1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.biochem.6b00090"},{"issue":"5","key":"2023090103250959000_2023.08.28.555139v1.2","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1021\/acs.biochem.6b01043","article-title":"Differences in Protein Concentration Dependence for Nucleation and Elongation in Light Chain Amyloid Formation","volume":"56","year":"2017","journal-title":"Biochemistry"},{"key":"2023090103250959000_2023.08.28.555139v1.3","doi-asserted-by":"publisher","DOI":"10.1080\/13506120802676781"},{"key":"2023090103250959000_2023.08.28.555139v1.4","first-page":"1","article-title":"Update, Novel Proteins, and Recommendations by the International Society of Amyloidosis (ISA) Nomenclature Committee","volume":"2022","author":"Amyloid Nomenclature","year":"2022","journal-title":"Amyloid"},{"key":"2023090103250959000_2023.08.28.555139v1.5","doi-asserted-by":"publisher","DOI":"10.1021\/bi7013773"},{"key":"2023090103250959000_2023.08.28.555139v1.6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1406050111"},{"key":"2023090103250959000_2023.08.28.555139v1.7","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0000981"},{"key":"2023090103250959000_2023.08.28.555139v1.8","doi-asserted-by":"publisher","DOI":"10.1002\/path.2607"},{"key":"2023090103250959000_2023.08.28.555139v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2009.11.005"},{"key":"2023090103250959000_2023.08.28.555139v1.10","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(99)09019-9"},{"key":"2023090103250959000_2023.08.28.555139v1.11","doi-asserted-by":"publisher","DOI":"10.1515\/hsz-2016-0178"},{"key":"2023090103250959000_2023.08.28.555139v1.12","doi-asserted-by":"publisher","DOI":"10.1006\/jmbi.2001.5001"},{"key":"2023090103250959000_2023.08.28.555139v1.13","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.91.12.5446"},{"key":"2023090103250959000_2023.08.28.555139v1.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-26845-0"},{"key":"2023090103250959000_2023.08.28.555139v1.15","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2016-10-743997"},{"key":"2023090103250959000_2023.08.28.555139v1.16","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M700545-MCP200"},{"key":"2023090103250959000_2023.08.28.555139v1.17","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1401564111"},{"key":"2023090103250959000_2023.08.28.555139v1.18","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2016.010"},{"issue":"1","key":"2023090103250959000_2023.08.28.555139v1.19","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1038\/s41572-018-0034-3","article-title":"Systemic Immunoglobulin Light Chain Amyloidosis","volume":"4","year":"2018","journal-title":"Nat Rev Dis Primers"},{"issue":"2","key":"2023090103250959000_2023.08.28.555139v1.20","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1080\/13506129.2021.1877129","article-title":"Early Events in Light Chain Aggregation at Physiological PH Reveal New Insights on Assembly, Stability, and Aggregate Dissociation","volume":"28","year":"2021","journal-title":"Amyloid"},{"issue":"29","key":"2023090103250959000_2023.08.28.555139v1.21","doi-asserted-by":"crossref","first-page":"3155","DOI":"10.1021\/acs.biochem.9b00311","article-title":"Mechanistic Insights into the Early Events in the Aggregation of Immunoglobulin Light Chains","volume":"58","year":"2019","journal-title":"Biochemistry"},{"key":"2023090103250959000_2023.08.28.555139v1.22","doi-asserted-by":"crossref","unstructured":"Morgan, G. 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