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Today,\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Y. pestis<\/jats:italic>\n                    <\/jats:styled-content>\n                    ,\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Y. enterocolitica<\/jats:italic>\n                    <\/jats:styled-content>\n                    , and\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Y. pseudotuberculosis<\/jats:italic>\n                    <\/jats:styled-content>\n                    remain clinically relevant human pathogens. Each of these pathogenic\n                    <jats:italic>Yersinia<\/jats:italic>\n                    species relies on a Type Three Secretion System (T3SS) for virulence, with the ATPase YscN playing a critical role in T3SS function. T3SS ATPases are responsible for powering apparatus formation and effector protein secretion through ATP hydrolysis. This study provides an extensive enzymatic characterization of recombinant YscN under several conditions, including variable pH and temperature, substrate and protein concentrations, and in the presence of putative inhibitors. Thermal stability data, assessed by circular dichroism, demonstrate that YscN exhibits increased stability in alkaline conditions, coinciding with greatest ATPase activity. Further, we report the first high\u2010resolution crystal structure of YscN and leverage homology data to model an oligomeric active site. Mutational analysis of a predicted active site residue confirms oligomerization as necessary for YscN ATPase activity and corroborates our oligomeric model and enzyme concentration\u2010dependent specific activity. Interestingly, however, AUC analysis reveals that the purified YscN predominantly exists as a monomer, despite oligomerization\u2010dependent active site formation. Thus, we propose that transient oligomeric interactions support the observed ATP hydrolysis. Together, these data uncover structural and environmental impacts on YscN activity that may support the highly specialized\n                    <jats:italic>Yersinia<\/jats:italic>\n                    pathogenic lifecycle and leverage its role in virulence in search of pan\u2010effective small molecule T3SS ATPase inhibitors.\n                  <\/jats:p>","DOI":"10.1002\/prot.70112","type":"journal-article","created":{"date-parts":[[2026,1,20]],"date-time":"2026-01-20T10:12:21Z","timestamp":1768903941000},"page":"1199-1211","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Structural and Biophysical Characterization of the\n                    <i>Yersinia<\/i>\n                    Type Three Secretion System\n                    <scp>ATPase YscN<\/scp>"],"prefix":"10.1002","volume":"94","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4607-1897","authenticated-orcid":false,"given":"Samuel 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USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicholas E.","family":"Dickenson","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry Utah State University  Logan Utah USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2026,1,20]]},"reference":[{"key":"e_1_2_11_2_1","unstructured":"World Health Organization \u201cNewsroom \u201d Plague 2022 https:\/\/www.who.int\/news\u2010room\/fact\u2010sheets\/detail\/plague."},{"key":"e_1_2_11_3_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0096513"},{"key":"e_1_2_11_4_1","unstructured":"Centers for Disease Control and Prevention \u201cPlague Clinical Care of Plague \u201d 2024 https:\/\/www.cdc.gov\/plague\/hcp\/clinical\u2010care\/index.html."},{"key":"e_1_2_11_5_1","first-page":"453","article-title":"Plague and Other Human Infections Caused by Yersinia Species","volume":"47","author":"Putzker 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