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| Targets |
SAMS targets AMP-activated protein kinase (AMPK), a key energy-sensing enzyme that plays a central role in cellular energy homeostasis. AMPK is activated in response to increases in cellular AMP/ATP ratio and phosphorylates downstream targets to regulate metabolism. The SAMS peptide is a specific substrate for AMPK based on the sequence around Ser79 on acetyl-CoA carboxylase, which is phosphorylated exclusively by AMPK. The peptide is not phosphorylated by any of five other protein kinases tested, demonstrating its specificity for AMPK. By serving as a substrate for AMPK, SAMS enables the measurement of AMPK activity in vitro.
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| ln Vitro |
SAMS peptide is a particular substrate of AMP-activated protein kinase (AMPK). The SAMS peptide was not phosphorylated by any of the five other pure protein kinases examined [1].
SAMS demonstrates potent in vitro activity as a specific substrate for AMPK. The peptide is based on the sequence around Ser79 on acetyl-CoA carboxylase, which is phosphorylated exclusively by AMPK. The SAMS peptide is not phosphorylated by any of five other protein kinases tested, demonstrating its specificity for AMPK. The peptide is more selective for AMPK than acetyl-CoA carboxylase itself and provides a convenient and sensitive assay for AMPK activity. The peptide contains a putative AMPK phosphorylation site (Ser568) of ChREBP and a PKA phosphorylation site (Thr666). Its activity is characterized by its ability to be phosphorylated by AMPK in kinase assays. |
| ln Vivo |
In vivo activity of SAMS is not applicable, as the peptide is used as a substrate for in vitro kinase assays rather than as a therapeutic agent. The peptide is not administered to animals for efficacy studies. Its primary use is in biochemical assays to measure AMPK activity in cell lysates or purified enzyme preparations. The peptide's specificity for AMPK makes it a valuable tool for studying AMPK function in various biological contexts.
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| Enzyme Assay |
In vitro kinase assays for SAMS involve measuring the phosphorylation of the peptide by AMPK. The assay typically uses recombinant AMPK or immunoprecipitated AMPK from cell lysates, incubated with the SAMS peptide, ATP (often with [gamma-32P]-ATP for radioactive detection), and appropriate buffer conditions. The phosphorylated peptide is separated from unreacted ATP by spotting onto P81 phosphocellulose paper or by using a filter-binding assay, and radioactivity is measured by liquid scintillation counting. Alternatively, non-radioactive methods using phospho-specific antibodies or mass spectrometry can be used. The peptide's specificity for AMPK is confirmed by testing against other kinases. The assay provides a convenient and sensitive method for measuring AMPK activity.
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| Cell Assay |
In vitro cellular assays using SAMS are typically performed to measure AMPK activity in cell lysates. Cells are treated with compounds that activate or inhibit AMPK, and lysates are prepared. The SAMS peptide is added to the lysates along with ATP, and AMPK activity is measured by quantifying phosphorylation of the peptide. This allows for the assessment of AMPK activity under various experimental conditions. The peptide's specificity for AMPK ensures that the measured activity reflects AMPK activity rather than other kinases. Cytotoxicity is not applicable for this substrate peptide. Results are expressed as AMPK activity (pmol phosphate incorporated per minute per mg protein) or as fold-change relative to control samples.
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| Animal Protocol |
In vivo animal studies are not applicable for SAMS, as the peptide is used as a substrate for in vitro kinase assays rather than as a therapeutic agent. The peptide is not administered to animals for efficacy studies. Its primary use is in biochemical assays to measure AMPK activity in tissue lysates from animal models.
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| Toxicity/Toxicokinetics |
Toxicology of SAMS is not applicable, as the peptide is used as a substrate for in vitro kinase assays rather than as a therapeutic agent. The peptide is not intended for in vivo administration. Standard laboratory safety precautions for handling peptides should be followed. Comprehensive toxicological characterization has not been reported, as the compound is not intended for human use. SAMS is strictly intended for research purposes as a biochemical tool.
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| References | |
| Additional Infomation |
SAMS (SAMS peptide) is a synthetic peptide substrate for AMP-activated protein kinase (AMPK) based on the sequence around Ser79 on acetyl-CoA carboxylase. It is more selective for AMPK than acetyl-CoA carboxylase itself and provides a convenient and sensitive assay for AMPK activity. The peptide has a molecular formula of C74H131N29O18S2 and a molecular weight of 1779.15 g/mol. SAMS is not phosphorylated by any of five other protein kinases tested. The peptide has not entered clinical trials and is available for research purposes only. SAMS is a valuable tool for studying AMPK activity and regulation.
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| Molecular Formula |
C74H131N29O18S2
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|---|---|
| Molecular Weight |
1779.14544
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| Exact Mass |
1777.97
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| CAS # |
125911-68-4
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| PubChem CID |
71773161
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| Appearance |
White to off-white solid powder
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| LogP |
2.372
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| Hydrogen Bond Donor Count |
27
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| Hydrogen Bond Acceptor Count |
27
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| Rotatable Bond Count |
62
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| Heavy Atom Count |
123
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| Complexity |
3490
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| Defined Atom Stereocenter Count |
14
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| SMILES |
C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CCSC)NC(=O)[C@H](CC2=CN=CN2)N
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| InChi Key |
YJAHUEAICBOBTC-CSVPVIMBSA-N
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| InChi Code |
InChI=1S/C74H131N29O18S2/c1-38(2)27-51(66(115)100-53(30-43-32-84-37-90-43)67(116)99-52(28-39(3)4)68(117)103-57(40(5)6)70(119)97-45(15-10-11-21-75)61(110)95-46(16-12-22-85-72(77)78)62(111)98-50(71(120)121)18-14-24-87-74(81)82)92-56(106)33-88-60(109)54(34-104)101-65(114)48(19-25-122-8)93-58(107)41(7)91-69(118)55(35-105)102-63(112)47(17-13-23-86-73(79)80)96-64(113)49(20-26-123-9)94-59(108)44(76)29-42-31-83-36-89-42/h31-32,36-41,44-55,57,104-105H,10-30,33-35,75-76H2,1-9H3,(H,83,89)(H,84,90)(H,88,109)(H,91,118)(H,92,106)(H,93,107)(H,94,108)(H,95,110)(H,96,113)(H,97,119)(H,98,111)(H,99,116)(H,100,115)(H,101,114)(H,102,112)(H,103,117)(H,120,121)(H4,77,78,85)(H4,79,80,86)(H4,81,82,87)/t41-,44-,45-,46-,47-,48-,49-,50-,51-,52-,53-,54-,55-,57-/m0/s1
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| Chemical Name |
(2S)-2-[[(2S)-2-[[(2S)-6-amino-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-amino-3-(1H-imidazol-5-yl)propanoyl]amino]-4-methylsulfanylbutanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-3-hydroxypropanoyl]amino]propanoyl]amino]-4-methylsulfanylbutanoyl]amino]-3-hydroxypropanoyl]amino]acetyl]amino]-4-methylpentanoyl]amino]-3-(1H-imidazol-5-yl)propanoyl]amino]-4-methylpentanoyl]amino]-3-methylbutanoyl]amino]hexanoyl]amino]-5-(diaminomethylideneamino)pentanoyl]amino]-5-(diaminomethylideneamino)pentanoic acid
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
H2O : ~100 mg/mL (~56.21 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (56.21 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.5621 mL | 2.8103 mL | 5.6207 mL | |
| 5 mM | 0.1124 mL | 0.5621 mL | 1.1241 mL | |
| 10 mM | 0.0562 mL | 0.2810 mL | 0.5621 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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