000118268 001__ 118268
000118268 005__ 20240319081008.0
000118268 0247_ $$2doi$$a10.1016/j.jbc.2022.102047
000118268 0248_ $$2sideral$$a129554
000118268 037__ $$aART-2022-129554
000118268 041__ $$aeng
000118268 100__ $$aBerardinelli, S.J.
000118268 245__ $$aO-fucosylation stabilizes the TSR3 motif in thrombospondin-1 by interacting with nearby amino acids and protecting a disulfide bond; 35597280
000118268 260__ $$c2022
000118268 5060_ $$aAccess copy available to the general public$$fUnrestricted
000118268 5203_ $$aThrombospondin type-1 repeats (TSRs) are small protein motifs containing six conserved cysteines forming three disulfide bonds that can be modified with an O-linked fucose. Protein O-fucosyltransferase 2 (POFUT2) catalyzes the addition of O-fucose to TSRs containing the appropriate consensus sequence, and the O-fucose modification can be elongated to a Glucose-Fucose disaccharide with the addition of glucose by ß3-glucosyltransferase (B3GLCT). Elimination of Pofut2 in mice results in embryonic lethality in mice, highlighting the biological significance of O-fucose modification on TSRs. Knockout of POFUT2 in HEK293T cells has been shown to cause complete or partial loss of secretion of many proteins containing O-fucosylated TSRs. In addition, POFUT2 is localized to the endoplasmic reticulum (ER) and only modifies folded TSRs, stabilizing their structures. These observations suggest that POFUT2 is involved in an ER quality control mechanism for TSR folding and that B3GLCT also participates in quality control by providing additional stabilization to TSRs. However, the mechanisms by which addition of these sugars result in stabilization are poorly understood. Here, we conducted molecular dynamics (MD) simulations and provide crystallographic and NMR evidence that the Glucose-Fucose disaccharide interacts with specific amino acids in the TSR3 domain in thrombospondin-1 that are within proximity to the O-fucosylation modification site resulting in protection of a nearby disulfide bond. We also show that mutation of these amino acids reduces the stabilizing effect of the sugars in vitro. These data provide mechanistic details regarding the importance of O-fucosylation and how it participates in quality control mechanisms inside the ER.
000118268 536__ $$9info:eu-repo/grantAgreement/ES/AEI/BFU2016-75633-P$$9info:eu-repo/grantAgreement/ES/AEI/PID2019-105451GB-I00$$9info:eu-repo/grantAgreement/ES/DGA/E34-17R$$9info:eu-repo/grantAgreement/ES/DGA/LMP58-18
000118268 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000118268 590__ $$a4.8$$b2022
000118268 592__ $$a1.805$$b2022
000118268 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b88 / 285 = 0.309$$c2022$$dQ2$$eT1
000118268 593__ $$aBiochemistry$$c2022$$dQ1
000118268 593__ $$aMolecular Biology$$c2022$$dQ1
000118268 593__ $$aCell Biology$$c2022$$dQ1
000118268 594__ $$a9.2$$b2022
000118268 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000118268 700__ $$aEletsky, A.
000118268 700__ $$0(orcid)0000-0002-4680-937X$$aValero-González, J.
000118268 700__ $$aIto, A.
000118268 700__ $$aManjunath, R.
000118268 700__ $$0(orcid)0000-0002-3122-9401$$aHurtado-Guerrero, R.
000118268 700__ $$aPrestegard, J.H.
000118268 700__ $$aWoods, R.J.
000118268 700__ $$aHaltiwanger, R.S.
000118268 773__ $$g298, 6 (2022), 102047 [11 pp.]$$pJ. biol. chem.$$tJournal of Biological Chemistry$$x0021-9258
000118268 8564_ $$s1914334$$uhttps://zaguan.unizar.es/record/118268/files/texto_completo.pdf$$yVersión publicada
000118268 8564_ $$s3482962$$uhttps://zaguan.unizar.es/record/118268/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000118268 909CO $$ooai:zaguan.unizar.es:118268$$particulos$$pdriver
000118268 951__ $$a2024-03-18-14:48:49
000118268 980__ $$aARTICLE