Structural basis of ligand binding to UDP-galactopyranose mutase from mycobacterium tuberculosis using substrate and tetrafluorinated substrate analogs
Structural basis of ligand binding to UDP-galactopyranose mutase from mycobacterium tuberculosis using substrate and tetrafluorinated substrate analogs
UDP-Galactopyranose mutase (UGM) is a flavin-containing enzyme that catalyses the reversible conversion of UDP-Galactopyranose (UDP-Galp) to UDP-Galactofuranose (UDP-Galf) and plays a key role in the biosynthesis of the mycobacterial cell wall galactofuran. A soluble, active form of UGM from Mycobacterium tuberculosis (MtUGM) was obtained from a dual His6-MBP tagged MtUGM construct. We present the first complex structures of MtUGM with bound substrate UDP-Galp (both oxidized flavin and reduced flavin). In addition, we have determined the complex structures of MtUGM with inhibitors (UDP and the dideoxy-tetrafluorinated analogs of both UDP-Galp (UDP-F4-Galp) and UDP-Galf (UDP-F4-Galf)), which represent the first complex structures of UGM with an analogue in the furanose form, as well as the first structures of dideoxy-tetrafluorinated sugar analogs bound to a protein. These structures provide detailed insight into ligand recognition by MtUGM and show a similar overall binding mode as reported for other prokaryotic UGMs. The binding of the ligand induces conformational changes in the enzyme, allowing ligand binding and active site closure. In addition, the complex structure of MtUGM with UDP-F4-Galf reveals the first detailed insight into how the furanose moiety binds to UGM. In particular, this study confirmed that the furanoside adopts a high energy conformation (4E) within the catalytic pocket. Moreover, these investigations provide structural insights to the enhanced binding of the dideoxy-tetrafluorinated sugars compared to unmodified analogs. These results will help in the design of carbohydrate mimetics and drug development, and show the enormous possibilities on the use of polyfluorination in the design of carbohydrate mimetics.
1230-1244
van Straaten, Karin E.
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A. Kuttiyatveetil, Jijin R.
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Sevrain, Charlotte M.
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Villaume, Sydney A.
10da5885-c936-4d10-a2d8-f6172409a551
Jimenez-Barbero, Jesus
84e4df1a-b6e9-4d7d-98b4-817fb9dc5e49
Linclau, Bruno
19b9cacd-b8e8-4c65-af36-6352cade84ba
Vincent, Stéphane P.
c1d1d147-b144-4e4c-9bf7-c1bc644587df
Sanders, David A.R.
d40a14de-5360-4191-a006-3d27c875747f
15 January 2015
van Straaten, Karin E.
372140f5-9410-4ccb-b2dc-d19ae2570efe
A. Kuttiyatveetil, Jijin R.
c9ffd6cd-f88e-4a95-85c7-24d62141be2d
Sevrain, Charlotte M.
5f06c1ae-b6f2-4634-95be-89bd998f9b8e
Villaume, Sydney A.
10da5885-c936-4d10-a2d8-f6172409a551
Jimenez-Barbero, Jesus
84e4df1a-b6e9-4d7d-98b4-817fb9dc5e49
Linclau, Bruno
19b9cacd-b8e8-4c65-af36-6352cade84ba
Vincent, Stéphane P.
c1d1d147-b144-4e4c-9bf7-c1bc644587df
Sanders, David A.R.
d40a14de-5360-4191-a006-3d27c875747f
van Straaten, Karin E., A. Kuttiyatveetil, Jijin R., Sevrain, Charlotte M., Villaume, Sydney A., Jimenez-Barbero, Jesus, Linclau, Bruno, Vincent, Stéphane P. and Sanders, David A.R.
(2015)
Structural basis of ligand binding to UDP-galactopyranose mutase from mycobacterium tuberculosis using substrate and tetrafluorinated substrate analogs.
Journal of the American Chemical Society, 137 (3), .
(doi:10.1021/ja511204p).
Abstract
UDP-Galactopyranose mutase (UGM) is a flavin-containing enzyme that catalyses the reversible conversion of UDP-Galactopyranose (UDP-Galp) to UDP-Galactofuranose (UDP-Galf) and plays a key role in the biosynthesis of the mycobacterial cell wall galactofuran. A soluble, active form of UGM from Mycobacterium tuberculosis (MtUGM) was obtained from a dual His6-MBP tagged MtUGM construct. We present the first complex structures of MtUGM with bound substrate UDP-Galp (both oxidized flavin and reduced flavin). In addition, we have determined the complex structures of MtUGM with inhibitors (UDP and the dideoxy-tetrafluorinated analogs of both UDP-Galp (UDP-F4-Galp) and UDP-Galf (UDP-F4-Galf)), which represent the first complex structures of UGM with an analogue in the furanose form, as well as the first structures of dideoxy-tetrafluorinated sugar analogs bound to a protein. These structures provide detailed insight into ligand recognition by MtUGM and show a similar overall binding mode as reported for other prokaryotic UGMs. The binding of the ligand induces conformational changes in the enzyme, allowing ligand binding and active site closure. In addition, the complex structure of MtUGM with UDP-F4-Galf reveals the first detailed insight into how the furanose moiety binds to UGM. In particular, this study confirmed that the furanoside adopts a high energy conformation (4E) within the catalytic pocket. Moreover, these investigations provide structural insights to the enhanced binding of the dideoxy-tetrafluorinated sugars compared to unmodified analogs. These results will help in the design of carbohydrate mimetics and drug development, and show the enormous possibilities on the use of polyfluorination in the design of carbohydrate mimetics.
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e-pub ahead of print date: 6 January 2015
Published date: 15 January 2015
Organisations:
Characterisation and Analytics
Identifiers
Local EPrints ID: 373298
URI: http://eprints.soton.ac.uk/id/eprint/373298
ISSN: 0002-7863
PURE UUID: 623ffcf0-c85d-4146-81c7-3fa51dc822e7
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Date deposited: 14 Jan 2015 16:41
Last modified: 15 Mar 2024 03:05
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Contributors
Author:
Karin E. van Straaten
Author:
Jijin R. A. Kuttiyatveetil
Author:
Charlotte M. Sevrain
Author:
Sydney A. Villaume
Author:
Jesus Jimenez-Barbero
Author:
Stéphane P. Vincent
Author:
David A.R. Sanders
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