Welcome to the BenchChem Online Store!
molecular formula C6H6N2O B132304 Nicotinamide-d4 CAS No. 347841-88-7

Nicotinamide-d4

Cat. No. B132304
M. Wt: 126.15 g/mol
InChI Key: DFPAKSUCGFBDDF-RHQRLBAQSA-N
Attention: For research use only. Not for human or veterinary use.
In Stock
  • Click on QUICK INQUIRY to receive a quote from our team of experts.
  • With the quality product at a COMPETITIVE price, you can focus more on your research.
Patent
US07737158B2

Procedure details

High performance liquid chromatography was used to detect Nampt reaction products. HPLC was performed with Waters 515 pumps and a 2487 detector (Waters, Mass.) with a Supelco LC-18-T column (15 cm×4.6 cm; Supelco, Pa.). The Nampt reaction was conducted at 37° C. for 15 min in 500 μl of reaction buffer (50 mM Tris-HCl [pH 7.5], 10 mM MgCl2, 50 mM nicotinamide, 0.2 mM PRPP) with 50 μg of the recombinant Nampt protein. The reaction was terminated by adding 125 μl of 1 M HClO4. Protein was then precipitated at 18,000 g, and 500 μl of the supernatant was neutralized with 40 μl of 3 M K2CO3. After centrifugation, 100 μl of sample was mixed with 400 μl of Buffer A (50 mM K2PO4/KHPO4, pH 7.0) and loaded into the HPLC system. The products from Nampt reaction were monitored by absorbance at 261 nm. Results of HPLC detection of Nampt reaction products showed that the mouse Nampt produced nicotinamide mononucleotide (NMN) from nicotinamide and PRPP (see, e.g., FIG. 4D). Nampt failed to catalyze the synthesis of nicotinic acid mononucleotide (NaMN) from nicotinic acid and PRPP (see, e.g., FIGS. 13A and 13B), confirming the substrate specificity of this enzyme. In isolated reactions, it was also confirmed that Nmnat catalyzed the synthesis of NAD from NMN and ATP.
Name
Tris-HCl
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Name
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Name
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One

Identifiers

REACTION_CXSMILES
C(O)C(N)(CO)CO.Cl.[Mg+2].[Cl-].[Cl-].[C:13]([NH2:21])(=[O:20])[C:14]1[CH:19]=[CH:18][CH:17]=[N:16][CH:15]=1.[CH2:22]([O:39][P:40]([OH:43])([OH:42])=[O:41])[C@H:23]1[O:27][C@H:26]([O:28][P:29]([O:32][P:33]([OH:36])([OH:35])=[O:34])([OH:31])=[O:30])[C@H:25]([OH:37])[C@@H:24]1[OH:38]>>[CH:18]1[CH:17]=[N+:16]([CH:26]2[O:27][C@H:23]([CH2:22][O:39][P:40]([O-:43])([OH:42])=[O:41])[C@@H:24]([OH:38])[C@H:25]2[OH:37])[CH:15]=[C:14]([C:13]([NH2:21])=[O:20])[CH:19]=1.[C:13]([NH2:21])(=[O:20])[C:14]1[CH:19]=[CH:18][CH:17]=[N:16][CH:15]=1.[CH2:22]([O:39][P:40]([OH:43])([OH:42])=[O:41])[C@H:23]1[O:27][C@H:26]([O:28][P:29]([O:32][P:33]([OH:36])([OH:35])=[O:34])([OH:31])=[O:30])[C@H:25]([OH:37])[C@@H:24]1[OH:38] |f:0.1,2.3.4|

Inputs

Step One
Name
Tris-HCl
Quantity
0 (± 1) mol
Type
reactant
Smiles
C(C(CO)(CO)N)O.Cl
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
[Mg+2].[Cl-].[Cl-]
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
C(C1=CN=CC=C1)(=O)N
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
C([C@@H]1[C@H]([C@H]([C@H](O1)OP(=O)(O)OP(=O)(O)O)O)O)OP(=O)(O)O

Conditions

Other
Conditions are dynamic
1
Details
See reaction.notes.procedure_details.

Workups

CUSTOM
Type
CUSTOM
Details
Nampt reaction products
CUSTOM
Type
CUSTOM
Details
The Nampt reaction
CUSTOM
Type
CUSTOM
Details
The reaction was terminated
ADDITION
Type
ADDITION
Details
by adding 125 μl of 1 M HClO4
CUSTOM
Type
CUSTOM
Details
Protein was then precipitated at 18,000 g, and 500 μl of the supernatant
ADDITION
Type
ADDITION
Details
After centrifugation, 100 μl of sample was mixed with 400 μl of Buffer A (50 mM K2PO4/KHPO4, pH 7.0)
CUSTOM
Type
CUSTOM
Details
The products from Nampt reaction
CUSTOM
Type
CUSTOM
Details
Results of HPLC detection of Nampt reaction products

Outcomes

Product
Name
Type
product
Smiles
C1=CC(=C[N+](=C1)C2[C@@H]([C@@H]([C@H](O2)COP(=O)(O)[O-])O)O)C(=O)N
Name
Type
product
Smiles
C(C1=CN=CC=C1)(=O)N
Name
Type
product
Smiles
C([C@@H]1[C@H]([C@H]([C@H](O1)OP(=O)(O)OP(=O)(O)O)O)O)OP(=O)(O)O

Source

Source
Open Reaction Database (ORD)
Description
The Open Reaction Database (ORD) is an open-access schema and infrastructure for structuring and sharing organic reaction data, including a centralized data repository. The ORD schema supports conventional and emerging technologies, from benchtop reactions to automated high-throughput experiments and flow chemistry. Our vision is that a consistent data representation and infrastructure to support data sharing will enable downstream applications that will greatly improve the state of the art with respect to computer-aided synthesis planning, reaction prediction, and other predictive chemistry tasks.
Patent
US07737158B2

Procedure details

High performance liquid chromatography was used to detect Nampt reaction products. HPLC was performed with Waters 515 pumps and a 2487 detector (Waters, Mass.) with a Supelco LC-18-T column (15 cm×4.6 cm; Supelco, Pa.). The Nampt reaction was conducted at 37° C. for 15 min in 500 μl of reaction buffer (50 mM Tris-HCl [pH 7.5], 10 mM MgCl2, 50 mM nicotinamide, 0.2 mM PRPP) with 50 μg of the recombinant Nampt protein. The reaction was terminated by adding 125 μl of 1 M HClO4. Protein was then precipitated at 18,000 g, and 500 μl of the supernatant was neutralized with 40 μl of 3 M K2CO3. After centrifugation, 100 μl of sample was mixed with 400 μl of Buffer A (50 mM K2PO4/KHPO4, pH 7.0) and loaded into the HPLC system. The products from Nampt reaction were monitored by absorbance at 261 nm. Results of HPLC detection of Nampt reaction products showed that the mouse Nampt produced nicotinamide mononucleotide (NMN) from nicotinamide and PRPP (see, e.g., FIG. 4D). Nampt failed to catalyze the synthesis of nicotinic acid mononucleotide (NaMN) from nicotinic acid and PRPP (see, e.g., FIGS. 13A and 13B), confirming the substrate specificity of this enzyme. In isolated reactions, it was also confirmed that Nmnat catalyzed the synthesis of NAD from NMN and ATP.
Name
Tris-HCl
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Name
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Name
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One

Identifiers

REACTION_CXSMILES
C(O)C(N)(CO)CO.Cl.[Mg+2].[Cl-].[Cl-].[C:13]([NH2:21])(=[O:20])[C:14]1[CH:19]=[CH:18][CH:17]=[N:16][CH:15]=1.[CH2:22]([O:39][P:40]([OH:43])([OH:42])=[O:41])[C@H:23]1[O:27][C@H:26]([O:28][P:29]([O:32][P:33]([OH:36])([OH:35])=[O:34])([OH:31])=[O:30])[C@H:25]([OH:37])[C@@H:24]1[OH:38]>>[CH:18]1[CH:17]=[N+:16]([CH:26]2[O:27][C@H:23]([CH2:22][O:39][P:40]([O-:43])([OH:42])=[O:41])[C@@H:24]([OH:38])[C@H:25]2[OH:37])[CH:15]=[C:14]([C:13]([NH2:21])=[O:20])[CH:19]=1.[C:13]([NH2:21])(=[O:20])[C:14]1[CH:19]=[CH:18][CH:17]=[N:16][CH:15]=1.[CH2:22]([O:39][P:40]([OH:43])([OH:42])=[O:41])[C@H:23]1[O:27][C@H:26]([O:28][P:29]([O:32][P:33]([OH:36])([OH:35])=[O:34])([OH:31])=[O:30])[C@H:25]([OH:37])[C@@H:24]1[OH:38] |f:0.1,2.3.4|

Inputs

Step One
Name
Tris-HCl
Quantity
0 (± 1) mol
Type
reactant
Smiles
C(C(CO)(CO)N)O.Cl
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
[Mg+2].[Cl-].[Cl-]
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
C(C1=CN=CC=C1)(=O)N
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
C([C@@H]1[C@H]([C@H]([C@H](O1)OP(=O)(O)OP(=O)(O)O)O)O)OP(=O)(O)O

Conditions

Other
Conditions are dynamic
1
Details
See reaction.notes.procedure_details.

Workups

CUSTOM
Type
CUSTOM
Details
Nampt reaction products
CUSTOM
Type
CUSTOM
Details
The Nampt reaction
CUSTOM
Type
CUSTOM
Details
The reaction was terminated
ADDITION
Type
ADDITION
Details
by adding 125 μl of 1 M HClO4
CUSTOM
Type
CUSTOM
Details
Protein was then precipitated at 18,000 g, and 500 μl of the supernatant
ADDITION
Type
ADDITION
Details
After centrifugation, 100 μl of sample was mixed with 400 μl of Buffer A (50 mM K2PO4/KHPO4, pH 7.0)
CUSTOM
Type
CUSTOM
Details
The products from Nampt reaction
CUSTOM
Type
CUSTOM
Details
Results of HPLC detection of Nampt reaction products

Outcomes

Product
Name
Type
product
Smiles
C1=CC(=C[N+](=C1)C2[C@@H]([C@@H]([C@H](O2)COP(=O)(O)[O-])O)O)C(=O)N
Name
Type
product
Smiles
C(C1=CN=CC=C1)(=O)N
Name
Type
product
Smiles
C([C@@H]1[C@H]([C@H]([C@H](O1)OP(=O)(O)OP(=O)(O)O)O)O)OP(=O)(O)O

Source

Source
Open Reaction Database (ORD)
Description
The Open Reaction Database (ORD) is an open-access schema and infrastructure for structuring and sharing organic reaction data, including a centralized data repository. The ORD schema supports conventional and emerging technologies, from benchtop reactions to automated high-throughput experiments and flow chemistry. Our vision is that a consistent data representation and infrastructure to support data sharing will enable downstream applications that will greatly improve the state of the art with respect to computer-aided synthesis planning, reaction prediction, and other predictive chemistry tasks.
Patent
US07737158B2

Procedure details

High performance liquid chromatography was used to detect Nampt reaction products. HPLC was performed with Waters 515 pumps and a 2487 detector (Waters, Mass.) with a Supelco LC-18-T column (15 cm×4.6 cm; Supelco, Pa.). The Nampt reaction was conducted at 37° C. for 15 min in 500 μl of reaction buffer (50 mM Tris-HCl [pH 7.5], 10 mM MgCl2, 50 mM nicotinamide, 0.2 mM PRPP) with 50 μg of the recombinant Nampt protein. The reaction was terminated by adding 125 μl of 1 M HClO4. Protein was then precipitated at 18,000 g, and 500 μl of the supernatant was neutralized with 40 μl of 3 M K2CO3. After centrifugation, 100 μl of sample was mixed with 400 μl of Buffer A (50 mM K2PO4/KHPO4, pH 7.0) and loaded into the HPLC system. The products from Nampt reaction were monitored by absorbance at 261 nm. Results of HPLC detection of Nampt reaction products showed that the mouse Nampt produced nicotinamide mononucleotide (NMN) from nicotinamide and PRPP (see, e.g., FIG. 4D). Nampt failed to catalyze the synthesis of nicotinic acid mononucleotide (NaMN) from nicotinic acid and PRPP (see, e.g., FIGS. 13A and 13B), confirming the substrate specificity of this enzyme. In isolated reactions, it was also confirmed that Nmnat catalyzed the synthesis of NAD from NMN and ATP.
Name
Tris-HCl
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Name
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One
Name
Quantity
0 (± 1) mol
Type
reactant
Reaction Step One

Identifiers

REACTION_CXSMILES
C(O)C(N)(CO)CO.Cl.[Mg+2].[Cl-].[Cl-].[C:13]([NH2:21])(=[O:20])[C:14]1[CH:19]=[CH:18][CH:17]=[N:16][CH:15]=1.[CH2:22]([O:39][P:40]([OH:43])([OH:42])=[O:41])[C@H:23]1[O:27][C@H:26]([O:28][P:29]([O:32][P:33]([OH:36])([OH:35])=[O:34])([OH:31])=[O:30])[C@H:25]([OH:37])[C@@H:24]1[OH:38]>>[CH:18]1[CH:17]=[N+:16]([CH:26]2[O:27][C@H:23]([CH2:22][O:39][P:40]([O-:43])([OH:42])=[O:41])[C@@H:24]([OH:38])[C@H:25]2[OH:37])[CH:15]=[C:14]([C:13]([NH2:21])=[O:20])[CH:19]=1.[C:13]([NH2:21])(=[O:20])[C:14]1[CH:19]=[CH:18][CH:17]=[N:16][CH:15]=1.[CH2:22]([O:39][P:40]([OH:43])([OH:42])=[O:41])[C@H:23]1[O:27][C@H:26]([O:28][P:29]([O:32][P:33]([OH:36])([OH:35])=[O:34])([OH:31])=[O:30])[C@H:25]([OH:37])[C@@H:24]1[OH:38] |f:0.1,2.3.4|

Inputs

Step One
Name
Tris-HCl
Quantity
0 (± 1) mol
Type
reactant
Smiles
C(C(CO)(CO)N)O.Cl
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
[Mg+2].[Cl-].[Cl-]
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
C(C1=CN=CC=C1)(=O)N
Name
Quantity
0 (± 1) mol
Type
reactant
Smiles
C([C@@H]1[C@H]([C@H]([C@H](O1)OP(=O)(O)OP(=O)(O)O)O)O)OP(=O)(O)O

Conditions

Other
Conditions are dynamic
1
Details
See reaction.notes.procedure_details.

Workups

CUSTOM
Type
CUSTOM
Details
Nampt reaction products
CUSTOM
Type
CUSTOM
Details
The Nampt reaction
CUSTOM
Type
CUSTOM
Details
The reaction was terminated
ADDITION
Type
ADDITION
Details
by adding 125 μl of 1 M HClO4
CUSTOM
Type
CUSTOM
Details
Protein was then precipitated at 18,000 g, and 500 μl of the supernatant
ADDITION
Type
ADDITION
Details
After centrifugation, 100 μl of sample was mixed with 400 μl of Buffer A (50 mM K2PO4/KHPO4, pH 7.0)
CUSTOM
Type
CUSTOM
Details
The products from Nampt reaction
CUSTOM
Type
CUSTOM
Details
Results of HPLC detection of Nampt reaction products

Outcomes

Product
Name
Type
product
Smiles
C1=CC(=C[N+](=C1)C2[C@@H]([C@@H]([C@H](O2)COP(=O)(O)[O-])O)O)C(=O)N
Name
Type
product
Smiles
C(C1=CN=CC=C1)(=O)N
Name
Type
product
Smiles
C([C@@H]1[C@H]([C@H]([C@H](O1)OP(=O)(O)OP(=O)(O)O)O)O)OP(=O)(O)O

Source

Source
Open Reaction Database (ORD)
Description
The Open Reaction Database (ORD) is an open-access schema and infrastructure for structuring and sharing organic reaction data, including a centralized data repository. The ORD schema supports conventional and emerging technologies, from benchtop reactions to automated high-throughput experiments and flow chemistry. Our vision is that a consistent data representation and infrastructure to support data sharing will enable downstream applications that will greatly improve the state of the art with respect to computer-aided synthesis planning, reaction prediction, and other predictive chemistry tasks.
© Copyright 2024 BenchChem. All Rights Reserved.