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Research Article
Steven J. Sucheck
University of Toledo
Corresponding Author
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Glycoside hydrolases (GH) are a large family of hydrolytic enzymes found in all domains of life. As such, they control a plethora of normal and pathogenic biological functions. Thus, understanding selective inhibition of GH enzymes at the atomic level can lead to the identification of new classes of therapeutics. In these studies, we identified a 4-⍺-glucoside of valienamine (8) as an inhibitor (IC50 of 484 ± 51 µM) of Streptomyces coelicolor (Sco) GlgE1-V279S which belongs to the GH13 Carbohydrate Active EnZyme (CAZy) family. The synthetic route to 8 and a closely related 4-⍺-glucoside of validamine (7) was achieved starting from readily available D-maltose. A key step in the synthesis was a chelation-controlled addition of vinylmagnesium bromide to a maltose-derived enone intermediate. X-ray structures of both 7 and 8 in complex with Sco GlgE1-V279S were solved to resolutions of 1.75 and 1.78 Å, respectively. Structural analysis revealed the valienamine derivative 8 binds the enzyme in an E2 conformation for the cyclohexene fragment. Also, the cyclohexene fragment shows a new hydrogen-bonding contact from the pseudo-diaxial C(3)-OH to the catalytic nucleophile Asp 394 at the enzyme active site. Asp 394, in fact, forms a bidentate interaction with both the C(3)-OH and C(7)-OH of the inhibitor. In contrast, compound 7 disrupts the catalytic sidechain interaction network of Sco GlgE1-V279S via steric interactions resulting in a conformation change in Asp 394. These findings will have implications for the design other aminocarbasugar-based GH13-inhibitors and will be useful for identifying more potent and selective inhibitors.
Keywords
Glycoside hydrolases (GH), cyclohexene fragment, aminocarbasugar-based GH13-inhibitors, hydrolytic enzymes
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No competing interests reported.
This is a list of supplementary files associated with this preprint. Click to download.
- Table1.tif
- Table2.tif
- Table3.tif
- Table4.tif
- Scheme1.png
Retrosynthetic strategy to prepare 4-α-glycoside derivatives of valienamine and validamine from maltose.
- Scheme2.png
Diastereoselective addition of vinylmagnesium bromide to enone 15.
- Scheme3.png
Synthesis of diastereomers 27A and 27B from D-(+)-maltose (20)
- Scheme4.png
RCM of dienes 27A and 27B.
- Scheme5.png
Synthesis of 4-α-glycoside derivatives of validamine 7 and valienamine 8 from alkene 28A
- supportinginfoaminosugarsfinal.docx
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CURRENT STATUS: Under Review
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Posted 22 Mar, 2021
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Editorial decision: Major revision
On 13 Apr, 2021
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On 19 Mar, 2021
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Editor invited
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This preprint is under consideration at Scientific Reports. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research Article
Steven J. Sucheck
University of Toledo
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 1
Posted 22 Mar, 2021
No community comments so far
Editorial decision: Major revision
On 13 Apr, 2021
Reviews received
Received 04 Apr, 2021
Reviewers agreed
On 19 Mar, 2021
Reviewers invited
Invitations sent on 17 Mar, 2021
Editor assigned
On 15 Mar, 2021
Editor invited
On 14 Mar, 2021
Submission checks complete
On 14 Mar, 2021
First submitted
On 12 Mar, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Glycoside hydrolases (GH) are a large family of hydrolytic enzymes found in all domains of life. As such, they control a plethora of normal and pathogenic biological functions. Thus, understanding selective inhibition of GH enzymes at the atomic level can lead to the identification of new classes of therapeutics. In these studies, we identified a 4-⍺-glucoside of valienamine (8) as an inhibitor (IC50 of 484 ± 51 µM) of Streptomyces coelicolor (Sco) GlgE1-V279S which belongs to the GH13 Carbohydrate Active EnZyme (CAZy) family. The synthetic route to 8 and a closely related 4-⍺-glucoside of validamine (7) was achieved starting from readily available D-maltose. A key step in the synthesis was a chelation-controlled addition of vinylmagnesium bromide to a maltose-derived enone intermediate. X-ray structures of both 7 and 8 in complex with Sco GlgE1-V279S were solved to resolutions of 1.75 and 1.78 Å, respectively. Structural analysis revealed the valienamine derivative 8 binds the enzyme in an E2 conformation for the cyclohexene fragment. Also, the cyclohexene fragment shows a new hydrogen-bonding contact from the pseudo-diaxial C(3)-OH to the catalytic nucleophile Asp 394 at the enzyme active site. Asp 394, in fact, forms a bidentate interaction with both the C(3)-OH and C(7)-OH of the inhibitor. In contrast, compound 7 disrupts the catalytic sidechain interaction network of Sco GlgE1-V279S via steric interactions resulting in a conformation change in Asp 394. These findings will have implications for the design other aminocarbasugar-based GH13-inhibitors and will be useful for identifying more potent and selective inhibitors.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10
No competing interests reported.
This is a list of supplementary files associated with this preprint. Click to download.
- Table1.tif
- Table2.tif
- Table3.tif
- Table4.tif
- Scheme1.png
Retrosynthetic strategy to prepare 4-α-glycoside derivatives of valienamine and validamine from maltose.
- Scheme2.png
Diastereoselective addition of vinylmagnesium bromide to enone 15.
- Scheme3.png
Synthesis of diastereomers 27A and 27B from D-(+)-maltose (20)
- Scheme4.png
RCM of dienes 27A and 27B.
- Scheme5.png
Synthesis of 4-α-glycoside derivatives of validamine 7 and valienamine 8 from alkene 28A
- supportinginfoaminosugarsfinal.docx
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