000151963 001__ 151963
000151963 005__ 20250321155444.0
000151963 0247_ $$2doi$$a10.1021/acscatal.1c01785
000151963 0248_ $$2sideral$$a126171
000151963 037__ $$aART-2021-126171
000151963 041__ $$aeng
000151963 100__ $$aPiniello B.
000151963 245__ $$aAsparagine Tautomerization in Glycosyltransferase Catalysis. The Molecular Mechanism of Protein O-Fucosyltransferase 1
000151963 260__ $$c2021
000151963 5060_ $$aAccess copy available to the general public$$fUnrestricted
000151963 5203_ $$aO-glycosylation is a post-translational protein modification essential to life. One of the enzymes involved in this process is protein O-fucosyltransferase 1 (POFUT1), which fucosylates threonine or serine residues within a specific sequence context of epidermal growth factor-like domains (EGF-LD). Unlike most inverting glycosyltransferases, POFUT1 lacks a basic residue in the active site that could act as a catalytic base to deprotonate the Thr/Ser residue of the EGF-LD acceptor during the chemical reaction. Using quantum mechanics/molecular mechanics (QM/MM) methods on recent crystal structures, as well as mutagenesis experiments, we uncover the enzyme catalytic mechanism, revealing that it involves proton shuttling through an active site asparagine, conserved among species, which undergoes tautomerization. This mechanism is consistent with experimental kinetic analysis of Caenorhabditis elegans POFUT1 Asn43 mutants, which ablate enzyme activity even if mutated to Asp, the canonical catalytic base in inverting glycosyltransferases. These results will aid inhibitor development for Notch-associated O-glycosylation disorders.
000151963 536__ $$9info:eu-repo/grantAgreement/ES/DGA-FEDER/E35-20R$$9info:eu-repo/grantAgreement/ES/DGA/LMP58-18$$9info:eu-repo/grantAgreement/ES/MICINN/IJCI-2017-32874$$9info:eu-repo/grantAgreement/ES/MICINN/PID2019-105451GB-I00
000151963 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000151963 590__ $$a13.7$$b2021
000151963 591__ $$aCHEMISTRY, PHYSICAL$$b19 / 165 = 0.115$$c2021$$dQ1$$eT1
000151963 592__ $$a4.202$$b2021
000151963 593__ $$aChemistry (miscellaneous)$$c2021$$dQ1
000151963 593__ $$aCatalysis$$c2021$$dQ1
000151963 594__ $$a20.8$$b2021
000151963 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000151963 700__ $$aLira-Navarrete E.$$uUniversidad de Zaragoza
000151963 700__ $$aTakeuchi H.
000151963 700__ $$aTakeuchi M.
000151963 700__ $$aHaltiwanger R.S.
000151963 700__ $$0(orcid)0000-0002-3122-9401$$aHurtado-Guerrero R.
000151963 700__ $$aRovira C.
000151963 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000151963 773__ $$g11, 15 (2021), 9926-9932$$pACS catal.$$tACS CATALYSIS$$x2155-5435
000151963 8564_ $$s3423364$$uhttps://zaguan.unizar.es/record/151963/files/texto_completo.pdf$$yVersión publicada
000151963 8564_ $$s2858992$$uhttps://zaguan.unizar.es/record/151963/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000151963 909CO $$ooai:zaguan.unizar.es:151963$$particulos$$pdriver
000151963 951__ $$a2025-03-21-14:40:44
000151963 980__ $$aARTICLE