{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,6]],"date-time":"2026-03-06T16:18:05Z","timestamp":1772813885732,"version":"3.50.1"},"reference-count":58,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2005,2,4]],"date-time":"2005-02-04T00:00:00Z","timestamp":1107475200000},"content-version":"vor","delay-in-days":4417,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cell Motil. Cytoskeleton"],"published-print":{"date-parts":[[1993,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>At least eight tropomyosin isoforms (hTM1, hTM2, hTM3, hTM4, hTM5, hTM5a, hTM5b, and hTMsm\u03b1) are expressed from four distinct genes in human fibroblasts. In order to elucidate isoform properties, we have subcloned hTM3 and hTM5 full\u2010length cDNAs, as well as their chimeric cDNAs into the bacterial expression pET8C system. Bacterially expressed tropomyosin isoforms (called PEThTM3, PEThTM5. PEThTM5\/3, and PEThTM3\/5) were purified and characterized. Under optimal binding conditions, the binding of PEThTM5 isoform to F\u2010actin was stronger than the PEThTM3 isoform. However, analysis of actin\u2010binding by the McGhee and von Hippel equation revealed that PEThTM3 exhibits higher cooperativity in binding than PEThTM5 does. Furthermore, the chimera PEThTM5\/3 which possessed the N\u2010terminal fragment of hTM5 fused to the C\u2010terminal fragment of hTM3 had even stronger actin binding ability. The reverse chimera PEThTM3\/5 which possessed the N\u2010terminal fragment of hTM3 fused to the C\u2010terminal fragment of hTM5 demonstrated greatly reduced affinity to actin filaments. In addition, both chimeras had different KCl requirements for optimal binding to F\u2010actin than their parental tropomyosins. A bacterially made C\u2010terminal fragment of human fibroblast caldesmon (PETCaD39) and native chicken gizzard caldesmon were both able to enhance the actin\u2010binding of these bacterially expressed tropomyosins. However, PETCaD39\u2032s enhancement of binding to F\u2010actin was greater for PEThTM5 than PEThTM3. Under 30 mM KCl and 4 mM MgCl<jats:sub>2<\/jats:sub>, the low M<jats:sub>r<\/jats:sub> isoform PEThTM5 appeared to be able to amplify the actin\u2010activated HMM ATPase activity by 4.7 fold, while the high M<jats:sub>r<\/jats:sub> isoform PEThTM3 stimulated the activity only 1.5 fold. The higher enhancement of ATPase activity by PEThTM5 than by PEThTM3 suggested that the low M<jats:sub>r<\/jats:sub> isoform hTM5 may be more involved in modulating nonmuscle cell motility than hTM3. These results further suggested that different isoforms of tropomyosin might have finite differences in their specific functions (e.g., cytoskeletal vs. motile) inside the cell. \u00a9 1993 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/cm.970260308","type":"journal-article","created":{"date-parts":[[2005,2,24]],"date-time":"2005-02-24T02:20:35Z","timestamp":1109211635000},"page":"248-261","source":"Crossref","is-referenced-by-count":59,"title":["In vitro functional characterization of bacterially expressed human fibroblast tropomyosin isoforms and their chimeric mutants"],"prefix":"10.1002","volume":"26","author":[{"given":"Robert E.","family":"Novy","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James R.","family":"Sellers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Li\u2010Fei","family":"Liu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jim Jung\u2010Ching","family":"Lin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2005,2,4]]},"reference":[{"key":"e_1_2_1_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1990.tb19478.x"},{"key":"e_1_2_1_3_1","doi-asserted-by":"crossref","first-page":"12873","DOI":"10.1016\/S0021-9258(18)90828-8","article-title":"Smooth muscle caldesmon: Rapid purification and F\u2010actin crosslinking properties","volume":"259","author":"Bretscher A.","year":"1984","journal-title":"J. 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