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A major limitation in the field consists currently in the lacking information on the signal(s) or chemoeffectors recognized by the majority of chemoreceptors, which in turn hampers understanding of how the environment has shaped chemotactic capabilities. A number of different approaches for signal identification have been reported that are often labour\u2010intensive. Here, we developed a straightforward NADH\u2010coupled CheA ATPase assay to systematically screen potential chemoeffectors. Obtained results are complemented with differential scanning fluorimetry and isothermal titration calorimetry analyses, as well as structural protein modelling to study the chemoreceptors of\n                    <jats:styled-content style=\"fixed-case\">\n                      <jats:italic>Pseudomonas parafulva<\/jats:italic>\n                    <\/jats:styled-content>\n                    PSR09\u201011288. Using this integrated approach, we identified L\u2010malate, citrate, L\u2010glutamine, and L\u2010asparagine as chemoattractants and revealed that chemoreceptor B2J77_03605 specifically recognizes L\u2010malate and citrate, whereas chemoreceptor B2J77_13170 binds L\u2010glutamine and L\u2010asparagine. The universality of this approach is shown by the fact that both chemoreceptors belong to different families and possess sensor domains that belong to the HBM and dCache families, respectively. This pipeline is a scalable workflow for chemoeffector identification, permitting systematic exploration of bacterial chemotactic capabilities.\n                  <\/jats:p>","DOI":"10.1111\/1462-2920.70408","type":"journal-article","created":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T10:37:53Z","timestamp":1788259073000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Rapid Screening of Bacterial Chemoeffectors of\n                    <scp>\n                      <i>Pseudomonas parafulva<\/i>\n                    <\/scp>\n                    Using a\n                    <scp>CheA ATPase<\/scp>\n                    Activity\u2010Based Assay"],"prefix":"10.1111","volume":"28","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4636-7951","authenticated-orcid":false,"given":"Rui","family":"Cui","sequence":"first","affiliation":[{"name":"State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences  Beijing P. R. China"},{"name":"College of Life Sciences, University of Chinese Academy of Sciences  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences  Beijing P. R. China"},{"name":"College of Life Sciences, University of Chinese Academy of Sciences  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ni\u2010Ye","family":"You","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences  Beijing P. R. China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9040-3166","authenticated-orcid":false,"given":"Tino","family":"Krell","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Environmental Protection Estaci\u00f3n Experimental del Zaid\u00edn Consejo Superior de Investigaciones Cient\u00edficas  Granada Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8683-019X","authenticated-orcid":false,"given":"De\u2010Feng","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Microbial Diversity and Innovative Utilization, Institute of Microbiology, Chinese Academy of Sciences  Beijing P. R. China"},{"name":"College of Life Sciences, University of Chinese Academy of Sciences  Beijing P. R. 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