{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T13:59:56Z","timestamp":1790258396141,"version":"4.1.0"},"reference-count":79,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2026,10]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Lactate dehydrogenases catalyze the reduction of pyruvate to lactate, the generation of which is of importance for the energetic metabolism of malignant cells. In particular, human cancer cells overexpress lactate dehydrogenase A (hLDH\u2010A), whose catalytic activity depends on its assembly into the corresponding homotetramer (denoted as hLDH\u20105), suggesting that compounds interfering with the protein\u2013protein interactions responsible for the generation of hLDH\u20105 should represent inhibitors of high selectivity. Not surprisingly, quite a number of competitive inhibitors were designed, synthesized, and tested against hLDH\u20105. However, only a few representatives of the repertoire accordingly obtained feature an appropriate selectivity. Here we show how the ionic strength and \u03b2\u2010NADH binding affect the monomers\u2010to\u2010tetramer hLDH\u2010A transition. Moreover, by taking advantage of the isolation of monomeric hLDH\u2010A at neutral pH, we tested the effectiveness of cyclic peptides and long\u2010chain dicarboxylates in hampering the assembly of hLDH\u20105. Interestingly, the cyclic peptide LCO15 and the long\u2010chain dicarboxylate crocetin were found to be very effective inhibitors of hLDH\u2010A: the catalytic activity of the enzyme was inhibited by 97% and 86% when these compounds were administered at 40 and 20\u2009\u03bcM, respectively. Furthermore, the amount of lactate produced by MCF7 human cancer cells was found to decrease by 30% in the presence of 80\u2009\u03bcM crocin (a diester of crocetin). Overall, our observations indicate that the assembly of hLDH\u2010A into catalytically competent hLDH\u20105 can be appropriately inhibited by means of cyclic peptides and long\u2010chain dicarboxylates.<\/jats:p>","DOI":"10.1002\/pro.70796","type":"journal-article","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T10:20:30Z","timestamp":1789726830000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The assembly of tetrameric human L\u2010lactate dehydrogenase is regulated by ionic strength, \u03b2\u2010\n                    <scp>NADH<\/scp>\n                    binding, cyclic peptides, and long\u2010chain dicarboxylates"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0391-7474","authenticated-orcid":false,"given":"Alessandra","family":"Stefan","sequence":"first","affiliation":[{"name":"Department of Pharmacy and Biotechnology University of Bologna  Bologna 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