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BiP also forms oligomers that influence its activity, particularly in binding polypeptide clients. Using single\u2010molecule approaches and mechanical and enzymatic activity assays, BiP monomer stability and oligomerization were analyzed both alone and in response to nucleotides and peptides. We find that the formation of BiP dimers exhibits biphasic behavior as a function of BiP concentration, suggesting concentration\u2010dependent changes in dimer dissociation constants. At low BiP concentrations, dimers were disrupted by peptide substrate and adenosine triphosphate (ATP) but remained unaffected by ADP or Adenosine 5\u2032\u2010O\u2010(3\u2010thio)triphosphate (ATP\u03b3S). At high concentrations, dimers are unaffected by peptides, but their assembly is inhibited by ATP and ATP\u03b3S to the same degree. These results suggest the formation of two distinct BiP dimers exhibiting unique binding affinities, kinetics, and potentially structures. We propose the existence of a high\u2010affinity dimer binding site within the SBD of BiP, while the formation of a low\u2010affinity dimer involves interactions between the lid and NBD of each protomer. Our findings demonstrate the importance of considering single\u2010molecule characteristics when interpreting bulk studies on protein function and regulation.<\/jats:p>","DOI":"10.1002\/pro.70690","type":"journal-article","created":{"date-parts":[[2026,7,9]],"date-time":"2026-07-09T10:56:21Z","timestamp":1783594581000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Immunoglobulin binding protein (\n                    <scp>BiP<\/scp>\n                    ) forms two types of dimers"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1939-4875","authenticated-orcid":false,"given":"Karina","family":"New","sequence":"first","affiliation":[{"name":"Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular, Facultad de Ciencias Qu\u00edmicas y Farmac\u00e9uticas Universidad de Chile  Santiago Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-4027-2702","authenticated-orcid":false,"given":"Miguel I. A.","family":"Lagos\u2010Espinoza","sequence":"additional","affiliation":[{"name":"Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular, Facultad de Ciencias Qu\u00edmicas y Farmac\u00e9uticas Universidad de Chile  Santiago Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9706-0952","authenticated-orcid":false,"given":"Nathalie","family":"Casanova\u2010Morales","sequence":"additional","affiliation":[{"name":"Facultad de Artes Liberales Universidad Adolfo Ib\u00e1\u00f1ez  Santiago Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5206-4378","authenticated-orcid":false,"given":"Roi","family":"Asor","sequence":"additional","affiliation":[{"name":"Physical and Theoretical Chemistry Laboratory, Department of Chemistry University of Oxford  Oxford UK"},{"name":"The Kavli Institute for Nanoscience Discovery  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3541-509X","authenticated-orcid":false,"given":"John William","family":"Young","sequence":"additional","affiliation":[{"name":"Physical and Theoretical Chemistry Laboratory, Department of Chemistry University of Oxford  Oxford UK"},{"name":"The Kavli Institute for Nanoscience Discovery  Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9760-174X","authenticated-orcid":false,"given":"Zahra","family":"Alavi","sequence":"additional","affiliation":[{"name":"Department of Physics Loyola Marymount University  Los Angeles California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6499-6957","authenticated-orcid":false,"given":"Christian A. 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