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dc.contributor.authorKeinänen, TA
dc.contributor.authorGrigorenko, N
dc.contributor.authorKhomutov, AR
dc.contributor.authorHuang, Q
dc.contributor.authorUimari, A
dc.contributor.authorAlhonen, L
dc.contributor.authorHyvönen, MT
dc.contributor.authorVepsäläinen, J
dc.date.accessioned2018-09-25T08:52:12Z
dc.date.available2018-09-25T08:52:12Z
dc.date.issued2018
dc.identifier.urihttps://erepo.uef.fi/handle/123456789/6960
dc.description.abstractEnzymes generally display strict stereospecificity and regioselectivity for their substrates. Here by using FAD-dependent human acetylpolyamine oxidase (APAO), human spermine (Spm) oxidase (SMOX) and yeast polyamine oxidase (Fms1), we demonstrate that these fundamental properties of the enzymes may be regulated using simple guide molecules, being either covalently attached to polyamines or used as a supplement to the substrate mixtures. APAO, which naturally metabolizes achiral N1-acetylated polyamines, displays aldehyde-controllable stereospecificity with chiral 1-methylated polyamines, like (R)- and (S)-1-methylspermidine (1,8-diamino-5-azanonane) (1-MeSpd). Among the novel N1-acyl derivatives of MeSpd, isonicotinic acid (P4) or benzoic acid (Bz) with (R)-MeSpd had Km of 3.6 ± 0.6/1.2 ± 0.7 µM and kcat of 5.2 ± 0.6/4.6 ± 0.7 s−1 respectively, while N1-AcSpd had Km 8.2 ± 0.4 µM and kcat 2.7 ± 0.0 s−1. On the contrary, corresponding (S)-MeSpd amides were practically inactive (kcat < 0.03 s−1) but they retained micromole level Km for APAO. SMOX did not metabolize any of the tested compounds (kcat < 0.05 s−1) that acted as non-competitive inhibitors having Ki ≥ 155 µM for SMOX. In addition, we tested (R,R)-1,12-bis-methylspermine (2,13-diamino-5,10-diazatetradecane) (R,R)-(Me2Spm) and (S,S)-Me2Spm as substrates for Fms1. Fms1 preferred (S,S)- to (R,R)-diastereoisomer, but with notably lower kcat in comparison with spermine. Interestingly, Fms1 was prone to aldehyde supplementation in its regioselectivity, i.e. the cleavage site of spermidine. Thus, aldehyde supplementation to generate aldimines or N-terminal substituents in polyamines, i.e. attachment of guide molecule, generates novel ligands with altered charge distribution changing the binding and catalytic properties with polyamine oxidases. This provides means for exploiting hidden capabilities of polyamine oxidases for controlling their regioselectivity and stereospecificity.
dc.language.isoenglanti
dc.publisherPortland Press Ltd.
dc.relation.ispartofseriesBioscience reports
dc.relation.urihttp://dx.doi.org/10.1042/BSR20180527
dc.rightsCC BY http://creativecommons.org/licenses/by/4.0/
dc.titleControlling the regioselectivity and stereospecificity of FAD-dependent polyamine oxidases with the use of amine-attached guide molecules as conformational modulators
dc.description.versionpublished version
dc.contributor.departmentSchool of Pharmacy, Activities
uef.solecris.id55902664en
dc.type.publicationTieteelliset aikakauslehtiartikkelit
dc.rights.accessrights© Authors
dc.relation.doi10.1042/BSR20180527
dc.description.reviewstatuspeerReviewed
dc.relation.articlenumberBSR20180527
dc.relation.issn0144-8463
dc.relation.issue4
dc.relation.volume38
dc.rights.accesslevelopenAccess
dc.type.okmA1
uef.solecris.openaccessOpen access -julkaisukanavassa ilmestynyt julkaisu


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