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The alternative exons of <jats:italic>Mhc<\/jats:italic> gene encode 4 distinct regions in the myosin motor domain, that is, the N\u2010terminal SH3\u2010like domain, one lip of the nucleotide\u2010binding pocket, the relay, and the converter. Here, we investigated the role of the alternative regions on the motor function of locust muscle myosin. Using Sf9\u2010baculovirus protein expression system, we expressed and purified 5 isoforms of the locust muscle myosin heavy meromyosin (HMM), including the major isoform in the thorax dorsal longitudinal flight muscle (FL1) and 4 isoforms expressed in the abdominal intersegmental muscle (AB1 to AB4). Among these 5 HMMs, FL1\u2010HMM displayed the highest level of actin\u2010activated adenosine triphosphatase (ATPase) activity (hereafter referred as ATPase activity). To identify the alternative region(s) responsible for the elevated ATPase activity of FL1\u2010HMM, we produced a number of chimeras of FL1\u2010HMM and AB4\u2010HMM. Substitution with the relay of AB4\u2010HMM (encoded by exon\u201014c) substantially decreased the ATPase activity of FL1\u2010HMM, and conversely, the relay of FL1\u2010HMM (encoded by exon\u201014a) enhanced the ATPase activity of AB4\u2010HMM. Mutagenesis showed that the exon\u201014a\u2010encoded residues Gly<jats:sup>474<\/jats:sup> and Asn<jats:sup>509<\/jats:sup> are responsible for the elevated ATPase activity of FL1\u2010HMM. Those results indicate that the alternative relay encoded by exon\u201014a\/c play a key role in regulating the ATPase activity of FL1\u2010HMM and AB4\u2010HMM.<\/jats:p>","DOI":"10.1111\/1744-7917.13257","type":"journal-article","created":{"date-parts":[[2023,7,25]],"date-time":"2023-07-25T06:37:06Z","timestamp":1690267026000},"page":"435-447","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Alternative relay regulates the adenosine triphosphatase activity of <i>Locusta migratoria<\/i> striated muscle myosin"],"prefix":"10.1111","volume":"31","author":[{"given":"Jie","family":"Hao","sequence":"first","affiliation":[{"name":"State Key Laboratory of Integrated Management of Insect Pests and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chang","family":"Liu","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Integrated Management of Insect Pests and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China"},{"name":"University of Chinese Academy of Sciences Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ning","family":"Zhang","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Integrated Management of Insect Pests and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Integrated Management of Insect Pests and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tong","family":"Ni","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Integrated Management of Insect Pests and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0397-1630","authenticated-orcid":false,"given":"Mingbo","family":"Qu","sequence":"additional","affiliation":[{"name":"School of Bioengeering Dalian University of Technology Dalian China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8677-9833","authenticated-orcid":false,"given":"Xiang\u2010dong","family":"Li","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Integrated Management of Insect Pests and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China"},{"name":"University of Chinese Academy of Sciences Beijing 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