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Their intrinsic resistance to antibiotics and resilience when exposed to environmental stress complicate treatment and underscore the need for discovering novel drug targets and developing new therapeutic strategies.\n                    <jats:italic>Mycobacterium hassiacum<\/jats:italic>\n                    , a thermophilic mycobacterium, is a promising source of stable proteins suitable for detailed structural analysis. In this study, we investigated its glucosyl\u20103\u2010phosphoglycerate synthase (GpgS), an enzyme that plays a key role in the biosynthesis of methylglucose lipopolysaccharides in\n                    <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                    and is implicated in responses to nitrogen starvation and thermal stress in different mycobacterial species. We confirmed the potential of\n                    <jats:italic>M. hassiacum<\/jats:italic>\n                    as a source of enzymes amenable to structural studies, as its GpgS ortholog yielded not only the first mycobacterial GpgS crystallographic structure at near\u2010atomic resolution (1.10\u2009\u00c5) but also multiple high\u2010resolution structures under different pH conditions and in complex with various substrates. These structural insights revealed a previously unrecognized aromatic binding pocket that is not only cryptic in nature but also capable of accommodating the substrates\n                    <jats:italic>p<\/jats:italic>\n                    \u2010aminobenzoic acid and\n                    <jats:italic>p<\/jats:italic>\n                    \u2010hydroxybenzaldehyde, which have been found to be relevant in its genetic context, thereby establishing a direct link between GpgS activity and broader metabolic pathways in mycobacteria. Our findings establish\n                    <jats:italic>M. hassiacum<\/jats:italic>\n                    as a valuable model for structural enzymology in mycobacteria and highlight GpgS as a potential drug target. The detailed structural data provided here offer a foundation for rational drug design, opening new avenues for the development of inhibitors targeting this key, potentially pleiotropic enzyme.\n                  <\/jats:p>","DOI":"10.1002\/pro.70713","type":"journal-article","created":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T08:31:56Z","timestamp":1784363516000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A cryptic aromatic binding pocket in the highly thermostable\n                    <scp>GpgS<\/scp>\n                    from\n                    <i>Mycobacterium hassiacum<\/i>\n                    hints at\n                    <scp>MGLP<\/scp>\n                    pathway crosstalk"],"prefix":"10.1002","volume":"35","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3398-9233","authenticated-orcid":false,"given":"Daniela","family":"Nunes\u2010Costa","sequence":"first","affiliation":[{"name":"CNC\u2010Center for Neuroscience and Cell Biology University of Coimbra  Coimbra Portugal"},{"name":"CIBB\u2010Centre for Innovative Biomedicine and Biotechnology University of Coimbra  Coimbra Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7604-792X","authenticated-orcid":false,"given":"Alexandra","family":"Silva","sequence":"additional","affiliation":[{"name":"i3S \u2010 Instituto de Investiga\u00e7\u00e3o e Inova\u00e7\u00e3o em Sa\u00fade Universidade do Porto  Porto Portugal"},{"name":"IBMC\u2010Instituto de Biologia Molecular e Celular Universidade do Porto  Porto 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