{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T12:30:49Z","timestamp":1787056249875,"version":"build-2736575974"},"reference-count":35,"publisher":"American Chemical Society (ACS)","issue":"16","license":[{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T00:00:00Z","timestamp":1785283200000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/na","name":"Department of Biochemistry, Western University","doi-asserted-by":"publisher","award":["R2276A10"],"award-info":[{"award-number":["R2276A10"]}],"id":[{"id":"10.13039\/na","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Tamavidin 2 is a biotin-binding protein of the avidin-like family, discovered in Tamogitake mushrooms, that has found a number of uses in biotechnology. Unlike avidin or streptavidin, it can be expressed in Escherichia coli as a soluble protein, making it a good partner in chimaeric proteins. Surprisingly, we found that the full-length protein purified from E. coli has a much lower affinity for biotin than that of avidin or streptavidin. In fact, a micromolar Kd was determined from fluorescence measurements, in contrast to the femtomolar Kd of avidin. Interestingly, treatments with trypsin or denaturing agents resulted in a large increase in the affinity. Modeling of the structure suggested that the C-terminal five amino acids obscured the biotin-binding site in the complete protein. Indeed, alterations of these five amino acids, either by truncation or individual mutation to alanine, resulted in a thousand-fold or greater increase in affinity for biotin. A truncated tamavidin was much more effective in forming complexes with biotinylated target proteins and as a green fluorescent protein (GFP) conjugate in binding assays. The inhibited state of the full-length protein and its activation by proteolysis suggest that tamavidin functions as a defensive protein to discourage predation.<\/jats:p>","DOI":"10.1021\/acs.biochem.6c00298","type":"journal-article","created":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T13:19:28Z","timestamp":1785849568000},"page":"2603-2610","source":"Crossref","is-referenced-by-count":0,"title":["Regulation of\nTamavidin\u2013Biotin Affinity by\nthe C-Terminal of the Native Protein"],"prefix":"10.1021","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2542-9156","authenticated-orcid":true,"given":"Eric H.","family":"Ball","sequence":"first","affiliation":[{"name":"Western University , , , ,","place":["London, Ontario, Canada, N6A 5C1"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Szabla","sequence":"additional","affiliation":[{"name":"Western University , , , ,","place":["London, Ontario, Canada, N6A 5C1"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gary","family":"Gill","sequence":"additional","affiliation":[{"name":"Western University , , , ,","place":["London, Ontario, Canada, N6A 5C1"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,29]]},"reference":[{"issue":"5","key":"2026081808122346600_cit1","doi-asserted-by":"publisher","first-page":"1383","DOI":"10.1111\/j.1742-4658.2009.06879.x","article-title":"Tamavidins--Novel Avidin-like\nBiotin-Binding Proteins from the Tamogitake Mushroom","volume":"276","author":"Takakura","year":"2009","journal-title":"FEBS J."},{"issue":"4","key":"2026081808122346600_cit2","doi-asserted-by":"publisher","first-page":"317","DOI":"10.1016\/j.jbiotec.2009.12.012","article-title":"Tamavidin, a Versatile Affinity Tag\nfor Protein Purification and Immobilization","volume":"145","author":"Takakura","year":"2010","journal-title":"J. Biotechnol."},{"key":"2026081808122346600_cit3","first-page":"5","article-title":"Introduction to\nAvidin-Biotin Technology","volume-title":"Methods in Enzymology","author":"Wilchek","year":"1990"},{"key":"2026081808122346600_cit4","doi-asserted-by":"publisher","first-page":"19.23.1","DOI":"10.1002\/0471140864.ps1923s74","article-title":"BioID: A Screen\nfor Protein-Protein\nInteractions","volume":"74","author":"Roux","year":"2013","journal-title":"Curr. Protoc. Protein Sci."},{"issue":"1","key":"2026081808122346600_cit5","doi-asserted-by":"publisher","first-page":"142","DOI":"10.1073\/pnas.87.1.142","article-title":"Expression of a\nCloned Streptavidin Gene in Escherichia Coli","volume":"87","author":"Sano","year":"1990","journal-title":"Proc. Natl.\nAcad. Sci. U.S.A."},{"issue":"2","key":"2026081808122346600_cit6","doi-asserted-by":"publisher","first-page":"192","DOI":"10.1006\/prep.1998.0930","article-title":"Production of a Soluble\nand Functional Recombinant Streptavidin inEscherichia Coli","volume":"14","author":"Gallizia","year":"1998","journal-title":"Protein Expression Purif."},{"key":"2026081808122346600_cit7","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1007\/978-1-59745-579-4_6","article-title":"High-Yield\nProduction and Purification of Recombinant T7-Tag Mature Streptavidin\nin Glucose-Stressed E. Coli","volume":"418","author":"Humbert","year":"2008","journal-title":"Methods Mol. Biol."},{"key":"2026081808122346600_cit8","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1016\/j.ymben.2016.12.013","article-title":"Biotin-Independent\nStrains of Escherichia Coli for Enhanced\nStreptavidin Production","volume":"40","author":"Jeschek","year":"2017","journal-title":"Metab. Eng."},{"issue":"2","key":"2026081808122346600_cit9","doi-asserted-by":"publisher","first-page":"571","DOI":"10.1016\/S0006-291X(05)80222-0","article-title":"Expression Vectors\nfor Streptavidin-Containing Chimeric\nProteins","volume":"176","author":"Sano","year":"1991","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"2026081808122346600_cit10","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1007\/978-1-4939-1034-2_8","article-title":"Expression, Purification, and Immobilization of Recombinant\nTamavidin 2 Fusion Proteins","volume":"1177","author":"Takakura","year":"2014","journal-title":"Methods Mol. Biol.."},{"issue":"4","key":"2026081808122346600_cit11","doi-asserted-by":"publisher","first-page":"390","DOI":"10.1016\/j.jbiosc.2015.08.016","article-title":"Immobilized\nSialyltransferase Fused to a Fungal Biotin-Binding Protein: Production,\nProperties, and Applications","volume":"121","author":"Kajiwara","year":"2016","journal-title":"J. Biosci. Bioeng."},{"issue":"4","key":"2026081808122346600_cit12","doi-asserted-by":"publisher","DOI":"10.1016\/j.xpro.2023.102633","article-title":"Immobilization of Lipid Nanorods onto Two-Dimensional\nCrystals of Protein Tamavidin 2 for High-Speed Atomic Force Microscopy","volume":"4","author":"Noshiro","year":"2023","journal-title":"STAR Protoc."},{"issue":"4","key":"2026081808122346600_cit13","doi-asserted-by":"publisher","DOI":"10.1002\/adbi.201900166","article-title":"Facile\nSynthesis\nand Characterization of a Novel Tamavidin-Luciferase Reporter Fusion\nProtein for Universal Signaling Applications","volume":"4","author":"Broyles","year":"2020","journal-title":"Adv. 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Biochem."},{"issue":"8016","key":"2026081808122346600_cit19","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate Structure\nPrediction of\nBiomolecular Interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"issue":"6","key":"2026081808122346600_cit20","doi-asserted-by":"publisher","first-page":"3031","DOI":"10.1021\/acs.jctc.7b00125","article-title":"The Rosetta All-Atom Energy Function\nfor Macromolecular Modeling and Design","volume":"13","author":"Alford","year":"2017","journal-title":"J. Chem.\nTheory Comput."},{"issue":"1","key":"2026081808122346600_cit21","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1002\/pro.2389","article-title":"Relaxation of Backbone Bond Geometry\nImproves Protein Energy Landscape\nModeling","volume":"23","author":"Conway","year":"2014","journal-title":"Protein Sci."},{"issue":"5259","key":"2026081808122346600_cit22","doi-asserted-by":"publisher","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage\nof Structural Proteins during the Assembly of the Head of Bacteriophage\nT4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"issue":"21","key":"2026081808122346600_cit23","doi-asserted-by":"publisher","first-page":"8537","DOI":"10.1021\/bi00447a040","article-title":"Shielding\nof Tryptophan Residues of Avidin by the Binding of Biotin","volume":"28","author":"Kurzban","year":"1989","journal-title":"Biochemistry"},{"issue":"12","key":"2026081808122346600_cit24","doi-asserted-by":"publisher","first-page":"2157","DOI":"10.1016\/j.str.2025.09.001","article-title":"Flexibility\nor Uncertainty? A Critical Assessment of AlphaFold 2 pLDDT","volume":"33","author":"Vander Meersche","year":"2025","journal-title":"Structure"},{"issue":"23","key":"2026081808122346600_cit25","doi-asserted-by":"publisher","first-page":"8485","DOI":"10.1128\/AEM.02286-12","article-title":"Biotin-Binding\nProteins in the Defense of Mushrooms against Predators and Parasites","volume":"78","author":"Bleuler-Martinez","year":"2012","journal-title":"Appl. Environ. Microbiol."},{"issue":"14","key":"2026081808122346600_cit26","doi-asserted-by":"publisher","first-page":"13250","DOI":"10.1074\/jbc.M414336200","article-title":"Novel Avidin-like\nProtein from a Root Nodule Symbiotic Bacterium, Bradyrhizobium Japonicum","volume":"280","author":"Nordlund","year":"2005","journal-title":"J. Biol. Chem."},{"issue":"5","key":"2026081808122346600_cit27","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0035962","article-title":"Structure of Bradavidin \u2013\nC-Terminal Residues\nAct as Intrinsic Ligands","volume":"7","author":"Leppiniemi","year":"2012","journal-title":"PLoS One"},{"issue":"2","key":"2026081808122346600_cit28","doi-asserted-by":"publisher","first-page":"139","DOI":"10.1016\/j.jsb.2015.06.020","article-title":"Hoefavidin:\nA Dimeric Bacterial Avidin with a C-Terminal Binding Tail","volume":"191","author":"Avraham","year":"2015","journal-title":"J. Struct. Biol."},{"issue":"3","key":"2026081808122346600_cit29","doi-asserted-by":"publisher","first-page":"738","DOI":"10.1016\/j.jmb.2005.11.086","article-title":"Crystallographic\nAnalysis of a Full-Length Streptavidin with Its C-Terminal Polypeptide\nBound in the Biotin Binding Site","volume":"356","author":"Le Trong","year":"2006","journal-title":"J. Mol. Biol."},{"issue":"29","key":"2026081808122346600_cit30","doi-asserted-by":"publisher","first-page":"13933","DOI":"10.1016\/S0021-9258(18)47884-2","article-title":"Characterization and Crystallization of Core Streptavidin","volume":"262","author":"Pa\u0308hler","year":"1987","journal-title":"J. Biol. Chem."},{"key":"2026081808122346600_cit31","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1016\/j.jbiotec.2016.01.032","article-title":"Constitutive\nProduction and Efficient Secretion of Soluble Full-Length Streptavidin\nby an Escherichia Coli \u201cLeaky\nMutant\u201d","volume":"221","author":"Mu\u0308ller","year":"2016","journal-title":"J. Biotechnol."},{"key":"2026081808122346600_cit32","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1385\/0-89603-481-X:119","article-title":"Expression and\nPurification of Recombinant Streptavidin-Containing Chimeric Proteins","volume-title":"Recombinant Protein Protocols","author":"Sano","year":"1997"},{"issue":"1","key":"2026081808122346600_cit33","doi-asserted-by":"publisher","first-page":"9","DOI":"10.1007\/s12033-011-9435-1","article-title":"Intercellular Production\nof Tamavidin 1, a Biotin-Binding Protein from Tamogitake Mushroom,\nConfers Resistance to the Blast Fungus Magnaporthe Oryzae in Transgenic\nRice","volume":"51","author":"Takakura","year":"2012","journal-title":"Mol. Biotechnol."},{"issue":"7","key":"2026081808122346600_cit34","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0069530","article-title":"Structure-Guided\nDesign of an Engineered Streptavidin\nwith Reusability to Purify Streptavidin-Binding Peptide Tagged Proteins\nor Biotinylated Proteins","volume":"8","author":"Wu","year":"2013","journal-title":"PLoS One"},{"issue":"1","key":"2026081808122346600_cit35","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1016\/j.jbiotec.2013.01.006","article-title":"Tamavidin\n2-REV: An Engineered Tamavidin\nwith Reversible Biotin-Binding Capability","volume":"164","author":"Takakura","year":"2013","journal-title":"J.\nBiotechnol."}],"container-title":["Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.biochem.6c00298","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/16\/2603\/66333971\/acs.biochem.6c00298.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/65\/16\/2603\/66333971\/acs.biochem.6c00298.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T12:12:39Z","timestamp":1787055159000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bichaw\/article\/65\/16\/2603\/5236972\/Regulation-of-Tamavidin-Biotin-Affinity-by-the-C"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,29]]},"references-count":35,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2026,7,29]]},"published-print":{"date-parts":[[2026,8,18]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.biochem.6c00298","relation":{},"ISSN":["0006-2960","1520-4995"],"issn-type":[{"value":"0006-2960","type":"print"},{"value":"1520-4995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,29]]}}}