Drug-dependent inhibition of nucleotide hydrolysis in the heterodimeric ABC multidrug transporter PatAB from <i>Streptococcus pneumoniae</i>

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dc.contributor.authorGuffick, Charlotte
dc.contributor.authorHsieh, Pei-Yu
dc.contributor.authorAli, Anam
dc.contributor.authorShi, Wilma
dc.contributor.authorHoward, Julie
dc.contributor.authorChinthapalli, Dinesh K.
dc.contributor.authorKong, Alex C.
dc.contributor.authorSalaa, Ihsene
dc.contributor.authorCrouch, Lucy, I
dc.contributor.authorAnsbro, Megan R.
dc.contributor.authorIsaacson, Shoshanna C.
dc.contributor.authorSingh, Himansha
dc.contributor.authorBarrera, Nelson P.
dc.contributor.authorNair, Asha, V
dc.contributor.authorRobinson, Carol, V
dc.contributor.authorDeery, Michael J.
dc.contributor.authorvan Veen, Hendrik W.
dc.date.accessioned2025-01-20T21:13:41Z
dc.date.available2025-01-20T21:13:41Z
dc.date.issued2022
dc.description.abstractThe bacterial heterodimeric ATP-binding cassette (ABC) multidrug exporter PatAB has a critical role in conferring antibiotic resistance in multidrug-resistant infections by Streptococcus pneumoniae. As with other heterodimeric ABC exporters, PatAB contains two transmembrane domains that form a drug translocation pathway for efflux and two nucleotide-binding domains that bind ATP, one of which is hydrolysed during transport. The structural and functional elements in heterodimeric ABC multidrug exporters that determine interactions with drugs and couple drug binding to nucleotide hydrolysis are not fully understood. Here, we used mass spectrometry techniques to determine the subunit stoichiometry in PatAB in our lactococcal expression system and investigate locations of drug binding using the fluorescent drug-mimetic azido-ethidium. Surprisingly, our analyses of azido-ethidium-labelled PatAB peptides point to ethidium binding in the PatA nucleotide-binding domain, with the azido moiety crosslinked to residue Q521 in the H-like loop of the degenerate nucleotide-binding site. Investigation into this compound and residue's role in nucleotide hydrolysis pointed to a reduction in the activity for a Q521A mutant and ethidium-dependent inhibition in both mutant and wild type. Most transported drugs did not stimulate or inhibit nucleotide hydrolysis of PatAB in detergent solution or lipidic nanodiscs. However, further examples for ethidium-like inhibition were found with propidium, novobiocin and coumermycin A1, which all inhibit nucleotide hydrolysis by a non-competitive mechanism. These data cast light on potential mechanisms by which drugs can regulate nucleotide hydrolysis by PatAB, which might involve a novel drug binding site near the nucleotide-binding domains.
dc.fuente.origenWOS
dc.identifier.doi10.1111/febs.16366
dc.identifier.eissn1742-4658
dc.identifier.issn1742-464X
dc.identifier.urihttps://doi.org/10.1111/febs.16366
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/93716
dc.identifier.wosidWOS:000754030100001
dc.issue.numero13
dc.language.isoen
dc.pagina.final3788
dc.pagina.inicio3770
dc.revistaFebs journal
dc.rightsacceso restringido
dc.subjectABC transporter
dc.subjectantibiotic resistance
dc.subjectdrug transport
dc.subjectnucleotide hydrolysis
dc.subjectstreptococcus
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleDrug-dependent inhibition of nucleotide hydrolysis in the heterodimeric ABC multidrug transporter PatAB from <i>Streptococcus pneumoniae</i>
dc.typeartículo
dc.volumen289
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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