Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts

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Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts. / de Las Rivas, Matilde; Coelho, Helena; Diniz, Ana; Lira-Navarrete, Erandi; Compañón, Ismael; Jiménez-Barbero, Jesús; T Schjoldager, Katrine; P Bennett, Eric; Y Vakhrushev, Sergey; Clausen, Henrik; Corzana, Francisco; Marcelo, Filipa; Hurtado-Guerrero, Ramon.

I: Chemistry: A European Journal, Bind 24, Nr. 33, 2018, s. 8382-8392.

Publikation: Bidrag til tidsskriftTidsskriftartikelfagfællebedømt

Harvard

de Las Rivas, M, Coelho, H, Diniz, A, Lira-Navarrete, E, Compañón, I, Jiménez-Barbero, J, T Schjoldager, K, P Bennett, E, Y Vakhrushev, S, Clausen, H, Corzana, F, Marcelo, F & Hurtado-Guerrero, R 2018, 'Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts', Chemistry: A European Journal, bind 24, nr. 33, s. 8382-8392. https://doi.org/10.1002/chem.201800701

APA

de Las Rivas, M., Coelho, H., Diniz, A., Lira-Navarrete, E., Compañón, I., Jiménez-Barbero, J., T Schjoldager, K., P Bennett, E., Y Vakhrushev, S., Clausen, H., Corzana, F., Marcelo, F., & Hurtado-Guerrero, R. (2018). Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts. Chemistry: A European Journal, 24(33), 8382-8392. https://doi.org/10.1002/chem.201800701

Vancouver

de Las Rivas M, Coelho H, Diniz A, Lira-Navarrete E, Compañón I, Jiménez-Barbero J o.a. Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts. Chemistry: A European Journal. 2018;24(33):8382-8392. https://doi.org/10.1002/chem.201800701

Author

de Las Rivas, Matilde ; Coelho, Helena ; Diniz, Ana ; Lira-Navarrete, Erandi ; Compañón, Ismael ; Jiménez-Barbero, Jesús ; T Schjoldager, Katrine ; P Bennett, Eric ; Y Vakhrushev, Sergey ; Clausen, Henrik ; Corzana, Francisco ; Marcelo, Filipa ; Hurtado-Guerrero, Ramon. / Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts. I: Chemistry: A European Journal. 2018 ; Bind 24, Nr. 33. s. 8382-8392.

Bibtex

@article{b3d79884353b45dd809e454166a3e5a0,
title = "Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts",
abstract = "The family of polypeptide 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 have been found to be related to different diseases. Recently, we have 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. Here, we have deciphered the molecular basis for F104S mutant inactivation. Saturation transfer difference NMR experiments demonstrate that the mutation induces loss of binding to peptide substrates. The analysis of the crystal structure of the F104S mutant bound to UDP-GalNAc, combined with molecular dynamics (MD) simulations, has revealed that the flexible loop is disordered and displays larger conformational changes in the mutant enzyme than in the wild-type (WT) enzyme. 19F-NMR experiments reveal that the WT enzyme reaches the active state only in the presence of UDP-GalNAc, providing compelling evidences that GalNAc-T2 adopts an UDP-GalNAc-dependent induced-fit mechanism. The F104S mutation precludes the enzyme to achieve the active conformation and concomitantly to bind peptide substrates. The present study provides new insights into the catalytic mechanism of the large family of GalNAc-Ts and how these enzymes orchestrate protein O-glycosylation.",
author = "{de Las Rivas}, Matilde and Helena Coelho and Ana Diniz and Erandi Lira-Navarrete and Ismael Compa{\~n}{\'o}n and Jes{\'u}s Jim{\'e}nez-Barbero and {T Schjoldager}, Katrine and {P Bennett}, Eric and {Y Vakhrushev}, Sergey and Henrik Clausen and Francisco Corzana and Filipa Marcelo and Ramon Hurtado-Guerrero",
note = "{\textcopyright} 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.",
year = "2018",
doi = "10.1002/chem.201800701",
language = "English",
volume = "24",
pages = "8382--8392",
journal = "Chemistry: A European Journal",
issn = "0947-6539",
publisher = "Wiley - V C H Verlag GmbH & Co. KGaA",
number = "33",

}

RIS

TY - JOUR

T1 - Structural analysis of a GalNAc-T2 mutant reveals an induced-fit catalytic mechanism for GalNAc-Ts

AU - de Las Rivas, Matilde

AU - Coelho, Helena

AU - Diniz, Ana

AU - Lira-Navarrete, Erandi

AU - Compañón, Ismael

AU - Jiménez-Barbero, Jesús

AU - T Schjoldager, Katrine

AU - P Bennett, Eric

AU - Y Vakhrushev, Sergey

AU - Clausen, Henrik

AU - Corzana, Francisco

AU - Marcelo, Filipa

AU - Hurtado-Guerrero, Ramon

N1 - © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

PY - 2018

Y1 - 2018

N2 - The family of polypeptide 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 have been found to be related to different diseases. Recently, we have 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. Here, we have deciphered the molecular basis for F104S mutant inactivation. Saturation transfer difference NMR experiments demonstrate that the mutation induces loss of binding to peptide substrates. The analysis of the crystal structure of the F104S mutant bound to UDP-GalNAc, combined with molecular dynamics (MD) simulations, has revealed that the flexible loop is disordered and displays larger conformational changes in the mutant enzyme than in the wild-type (WT) enzyme. 19F-NMR experiments reveal that the WT enzyme reaches the active state only in the presence of UDP-GalNAc, providing compelling evidences that GalNAc-T2 adopts an UDP-GalNAc-dependent induced-fit mechanism. The F104S mutation precludes the enzyme to achieve the active conformation and concomitantly to bind peptide substrates. The present study provides new insights into the catalytic mechanism of the large family of GalNAc-Ts and how these enzymes orchestrate protein O-glycosylation.

AB - The family of polypeptide 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 have been found to be related to different diseases. Recently, we have 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. Here, we have deciphered the molecular basis for F104S mutant inactivation. Saturation transfer difference NMR experiments demonstrate that the mutation induces loss of binding to peptide substrates. The analysis of the crystal structure of the F104S mutant bound to UDP-GalNAc, combined with molecular dynamics (MD) simulations, has revealed that the flexible loop is disordered and displays larger conformational changes in the mutant enzyme than in the wild-type (WT) enzyme. 19F-NMR experiments reveal that the WT enzyme reaches the active state only in the presence of UDP-GalNAc, providing compelling evidences that GalNAc-T2 adopts an UDP-GalNAc-dependent induced-fit mechanism. The F104S mutation precludes the enzyme to achieve the active conformation and concomitantly to bind peptide substrates. The present study provides new insights into the catalytic mechanism of the large family of GalNAc-Ts and how these enzymes orchestrate protein O-glycosylation.

U2 - 10.1002/chem.201800701

DO - 10.1002/chem.201800701

M3 - Journal article

C2 - 29601100

VL - 24

SP - 8382

EP - 8392

JO - Chemistry: A European Journal

JF - Chemistry: A European Journal

SN - 0947-6539

IS - 33

ER -

ID: 196436413