000078766 001__ 78766
000078766 005__ 20200117211600.0
000078766 0247_ $$2doi$$a10.1002/chem.201800701
000078766 0248_ $$2sideral$$a107167
000078766 037__ $$aART-2018-107167
000078766 041__ $$aeng
000078766 100__ $$0(orcid)0000-0002-1924-334X$$ade las Rivas, M.$$uUniversidad de Zaragoza
000078766 245__ $$aStructural Analysis of a GalNAc-T2 Mutant Reveals an Induced-Fit Catalytic Mechanism for GalNAc-Ts
000078766 260__ $$c2018
000078766 5060_ $$aAccess copy available to the general public$$fUnrestricted
000078766 5203_ $$aThe family of polypeptide N-acetylgalactosamine (GalNAc) transferases (GalNAc-Ts) orchestrates the initiating step of mucin-type protein O-glycosylation by transfer of GalNAc moieties to serine and threonine residues in proteins. Deficiencies and dysregulation of GalNAc-T isoenzymes are related to different diseases. Recently, it has been demonstrated that an inactive GalNAc-T2 mutant (F104S), which is not located at the active site, induces low levels of high-density lipoprotein cholesterol (HDL-C) in humans. Herein, the molecular basis for F104S mutant inactivation has been deciphered. Saturation transfer difference NMR spectroscopy experiments demonstrate that the mutation induces loss of binding to peptide substrates. Analysis of the crystal structure of the F104S mutant bound to UDP-GalNAc (UDP=uridine diphosphate), combined with molecular dynamics (MD) simulations, has revealed that the flexible loop is disordered and displays larger conformational changes in the mutant enzyme than that in the wild-type (WT) enzyme. 19F NMR spectroscopy experiments reveal that the WT enzyme only reaches the active state in the presence of UDP-GalNAc, which provides compelling evidence that GalNAc-T2 adopts a UDP-GalNAc-dependent induced-fit mechanism. The F104S mutation precludes the enzyme from achieving the active conformation and concomitantly binding peptide substrates. This study provides new insights into the catalytic mechanism of the large family of GalNAc-Ts and how these enzymes orchestrate protein O-glycosylation.
000078766 536__ $$9info:eu-repo/grantAgreement/ES/MINECO/CTQ2015-67727-R$$9info:eu-repo/grantAgreement/ES/MINECO/CTQ2015-64597-C2-1-P$$9info:eu-repo/grantAgreement/ES/MINECO/CTQ2013-44367-C2-2-P$$9info:eu-repo/grantAgreement/ES/MINECO/BFU2016-75633-P$$9info:eu-repo/grantAgreement/EC/FP7/609409/EU/Supporting transnational mobility within the European life sciences by cofunding of the EMBO Fellowship Programme/LTFCOFUND2013$$9info:eu-repo/grantAgreement/EC/FP7/283570/EU/Transnational access and enhancement of integrated Biological Structure determination at synchrotron X-ray radiation facilities/BIOSTRUCT-X
000078766 540__ $$9info:eu-repo/semantics/openAccess$$aAll rights reserved$$uhttp://www.europeana.eu/rights/rr-f/
000078766 590__ $$a5.16$$b2018
000078766 591__ $$aCHEMISTRY, MULTIDISCIPLINARY$$b36 / 172 = 0.209$$c2018$$dQ1$$eT1
000078766 592__ $$a1.842$$b2018
000078766 593__ $$aCatalysis$$c2018$$dQ1
000078766 593__ $$aOrganic Chemistry$$c2018$$dQ1
000078766 593__ $$aChemistry (miscellaneous)$$c2018$$dQ1
000078766 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/acceptedVersion
000078766 700__ $$aCoelho, H.
000078766 700__ $$aDiniz, A.
000078766 700__ $$aLira-Navarrete, E.
000078766 700__ $$aCompañón, I.
000078766 700__ $$aJiménez-Barbero, J.
000078766 700__ $$aSchjoldager, K.T.
000078766 700__ $$aBennett, E.P.
000078766 700__ $$aVakhrushev, S.Y.
000078766 700__ $$aClausen, H.
000078766 700__ $$aCorzana, F.
000078766 700__ $$aMarcelo, F.
000078766 700__ $$0(orcid)0000-0002-3122-9401$$aHurtado-Guerrero, R.$$uUniversidad de Zaragoza
000078766 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000078766 773__ $$g24, 33 (2018), 8382-8392$$pChemistry (Weinh.)$$tChemistry - A European Journal$$x0947-6539
000078766 8564_ $$s1216316$$uhttps://zaguan.unizar.es/record/78766/files/texto_completo.pdf$$yPostprint
000078766 8564_ $$s31984$$uhttps://zaguan.unizar.es/record/78766/files/texto_completo.jpg?subformat=icon$$xicon$$yPostprint
000078766 909CO $$ooai:zaguan.unizar.es:78766$$particulos$$pdriver
000078766 951__ $$a2020-01-17-21:11:05
000078766 980__ $$aARTICLE