MTSEA
(Synonyms: 2-Aminoethyl methanethiosulfonate, Methanethiosulfonate Ethylammonium) 目录号 : GC44252
MTSEA 是一种半胱氨酸修饰剂，半胱氨酸'的巯基侧链(-SH)可与MTSEA试剂中的硫形成共价二硫键.MTSEA 用于证明在人多巴胺 D2 受体的结合位点缝隙中存在水可接近的半胱氨酸。
Cas No.:16599-33-0
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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- Purity: >95.00%
- COA (Certificate Of Analysis)
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- Datasheet
MTSEA [(2-aminoethyl) methanethiosulfonate] is a cysteine modifying agent [1], cysteine's sulfhydral side chain (-SH) can form a covalent disulfide bond with sulfur in the MTSEA reagent [2].
MTSEA was used to demonstrate the presence of a water-accessible cysteine within the binding-site crevice of the human dopamine D2 receptor [3]. MTSEA was demonstrated rotation of TM6 (upon receptor activation) by the appearance of an MTSEA-accessible cysteine in a constitutively active mutant of the β2-adrenergic receptor not accessible in the wild-type receptor [4]. The effect of MTSEA on ligand binding, in conjunction with site-directed mutagenesis, was used to define intramolecular contacts in the neurokinin-2 tachykinin receptor [5].
References:
[1]. Choi YB, Tenneti L, Le DA, Ortiz J, Bai G, Chen HS, Lipton SA. Molecular basis of NMDA receptor-coupled ion channel modulation by S-nitrosylation. Nature neuroscience. 2000 Jan;3(1):15-21.
[2]. O'Reilly JP, Shockett PE. Time-and state-dependent effects of methanethiosulfonate ethylammonium (MTSEA) exposure differ between heart and skeletal muscle voltage-gated Na+ channels. Biochimica et Biophysica Acta (BBA)-Biomembranes. 2012 Mar 1;1818(3):443-7.
[3]. Javitch JA, Li X, Kaback J, Karlin A. A cysteine residue in the third membrane-spanning segment of the human D2 dopamine receptor is exposed in the binding-site crevice. Proceedings of the National Academy of Sciences. 1994 Oct 25;91(22):10355-9.
[4]. Rasmussen SG, Jensen AD, Liapakis G, Ghanouni P, Javitch JA, Gether U. Mutation of a highly conserved aspartic acid in the β2 adrenergic receptor: constitutive activation, structural instability, and conformational rearrangement of transmembrane segment 6. Molecular pharmacology. 1999 Jul 1;56(1):175-84.
[5]. Bhogal N, Blaney FE, Ingley PM, Rees J, Findlay JB. Evidence for the proximity of the extreme N-terminus of the neurokinin-2 (NK2) tachykinin receptor to cys167 in the putative fourth transmembrane helix. Biochemistry. 2004 Mar 23;43(11):3027-38.
MTSEA [(2-aminoethyl) methanethiosulfonate] 是一种半胱氨酸修饰剂[1]，半胱氨酸'的巯基侧链(-SH)可与MTSEA试剂中的硫形成共价二硫键[2].
MTSEA 用于证明在人多巴胺 D2 受体的结合位点缝隙中存在水可接近的半胱氨酸[3]。通过在 β 的组成型活性突变体中出现 MTSEA 可接近的半胱氨酸，证明了 MTSEA TM6 的旋转（在受体激活后）；2-肾上腺素能受体在野生型受体中不可接近[4]。MTSEA 对配体结合的影响，结合定点诱变，被用来定义神经激肽-2 速激肽受体的分子内接触[5]。
| Cas No. | 16599-33-0 | SDF | |
| 别名 | 2-Aminoethyl methanethiosulfonate, Methanethiosulfonate Ethylammonium | ||
| Canonical SMILES | NCCSS(C)(=O)=O.Br | ||
| 分子式 | C3H9NO2S2•HBr | 分子量 | 236.1 |
| 溶解度 | DMF: 30 mg/ml,DMSO: 30 mg/ml,Ethanol: 30 mg/ml,PBS (pH 7.2): 10 mg/ml | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 4.2355 mL | 21.1775 mL | 42.3549 mL |
| 5 mM | 0.8471 mL | 4.2355 mL | 8.471 mL |
| 10 mM | 0.4235 mL | 2.1177 mL | 4.2355 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
MTSEA prevents ligand binding to the human melanocortin-4 receptor by modification of cysteine 130 in transmembrane helix 3
FEBS Lett 2005 Jan 3;579(1):285-91.PMID:15620728DOI:10.1016/j.febslet.2004.11.087.
We have investigated the effect of the sulfhydryl-reactive reagent, methyl thiosulfonate ethylammonium (MTSEA), on ligand binding to the human melanocortin-4 (MC4) receptor stably expressed in HEK-293 cells. MTSEA inhibited binding of the agonist, 125I-NDPalpha-MSH, and the antagonist, 125I-SHU9119, in a concentration-dependent manner. Pre-incubation of cells with either the agonist or antagonist protected from subsequent MTSEA inhibition of radioligand binding. Mutation of Cys130 in transmembrane helix 3 to alanine, whilst not affecting ligand binding, led to a complete loss of the inhibitory effect of MTSEA. Since other types of sulfhydryl-reactive reagents had no effect on ligand binding, we conclude that covalent modification of Cys130 by MTSEA disrupts ligand binding by neutralising a close-by negative charge, most likely on Asp126.
Verapamil- and state-dependent effect of 2-aminoethylmethanethiosulphonate (MTSEA) on hK(v)1.3 channels
Br J Pharmacol 2012 Nov;167(6):1378-88.PMID:22748056DOI:10.1111/j.1476-5381.2012.02092.x.
Background and purpose: T-cells usually express voltage-gated K(v) 1.3 channels. These channels are distinguished by their typical C-type inactivation. Therefore, to be able to rationally design drugs specific for the C-type inactivation state that may have therapeutic value in autoimmune disease therapy, it is necessary to identify those amino acids that are accessible for drug binding in C-type inactivated channels. Experimental approach: The influence of 2-aminoethylmethanethiosulphonate (MTSEA) on currents through wild-type human K(v)1.3 (hK(v)1.3) and three mutant channels, hK(v)1.3_L418C, hK(v)1.3_T419C and hK(v)1.3_I420C, in the closed, open and inactivated states was investigated by the patch-clamp technique. Key results: Currents through hK(v)1.3_L418C and hK(v)1.3_T419C channels were irreversibly reduced after the external application of MTSEA in the open state but not in the inactivated and closed states. Currents through hK(v)1.3_I420C channels were irreversibly reduced in the open and inactivated states but not in the closed state. In the presence of verapamil, the MTSEA modification of hK(v)1.3_T419C and hK(v)1.3_I420C channels was prevented, while the MTSEA modification of hK(v)1.3_L418C channels was unaffected. Conclusion and implications: From our experiments, we conclude that the activation gate of all mutant channels must be open for modification by MTSEA and must also be open during inactivation. In addition, the relative movement of the S6 segments that occur during C-type inactivation includes a movement of the side chains of the amino acids at positions 418 and 419 away from the pore lining. Furthermore, the overlapping binding site for MTSEA and verapamil does not include position 418 in hK(v) 1.3 channels.
Time- and state-dependent effects of methanethiosulfonate ethylammonium (MTSEA) exposure differ between heart and skeletal muscle voltage-gated Na(+) channels
Biochim Biophys Acta 2012 Mar;1818(3):443-7.PMID:22155680DOI:10.1016/j.bbamem.2011.11.031.
The substituted-cysteine scanning method (SCAM) is used to study conformational changes in proteins. Experiments using SCAM involve site-directed mutagenesis to replace native amino acids with cysteine and subsequent exposure to a methanethiosulfonate (MTS) reagent such as methanethiosulfonate ethylammonium (MTSEA). These reagents react with substituted-cysteines and can provide functional information about relative positions of amino acids within a protein. In the human heart voltage-gated Na(+) channel hNav1.5 there is a native cysteine at position C373 that reacts rapidly with MTS reagents resulting in a large reduction in whole-cell Na(+) current (I(Na)). Therefore, in order to use SCAM in studies in this isoform, this native cysteine is mutated to a non-reactive residue, e.g., tyrosine. This mutant, hNav1.5-C373Y, is resistant to the MTS-mediated decrease in I(Na). Here we show that this resistance is time- and state-dependent. With relatively short exposure times to MTSEA (<4min), there is little effect on I(Na). However, with longer exposures (4-8min), there is a large decrease in I(Na), but this effect is only found when hNav1.5-C373Y is inactivated (fast or slow) - MTSEA has little effect in the closed state. Additionally, this long-term, state-dependent effect is not seen in human skeletal muscle Na(+) channel isoform hNav1.4, which has a native tyrosine at the homologous site C407. We conclude that differences in molecular determinants of inactivation between hNav1.4 and hNav1.5 underlie the difference in response to MTSEA exposure.
MTSEA potentiates 5-HT3 receptors containing the nicotinic alpha4 subunit
Neuropharmacology 1999 Dec;38(12):1913-5.PMID:10608286DOI:10.1016/s0028-3908(99)00109-4.
In order to study the subunit composition of 5-HT3 receptors (5-HT3R), we report that (2-aminoethyl)methanethiosulfonate (MTSEA) can enhance the function of both nicotinic ACh receptors (nAChRs) comprised of alpha4/beta2 subunits, and heteromeric channels assembled from serotonin 5-HT3R and alpha4 nAChR subunits. MTSEA has no effect on homomeric 5-HT3 receptor channels.
A selective and sensitive detection system for 4-thiouridine modification in RNA
RNA 2023 Feb;29(2):241-251.PMID:PMC9891261DOI:10.1261/rna.079445.122.
4-Thiouridine (s4U) is a modified nucleoside, found at positions 8 and 9 in tRNA from eubacteria and archaea. Studies of the biosynthetic pathway and physiological role of s4U in tRNA are ongoing in the tRNA modification field. s4U has also recently been utilized as a biotechnological tool for analysis of RNAs. Therefore, a selective and sensitive system for the detection of s4U is essential for progress in the fields of RNA technologies and tRNA modification. Here, we report the use of biotin-coupled 2-aminoethyl-methanethiosulfonate (MTSEA biotin-XX) for labeling of s4U and demonstrate that the system is sensitive and quantitative. This technique can be used without denaturation; however, addition of a denaturation step improves the limit of detection. Thermus thermophilus tRNAs, which abundantly contain 5-methyl-2-thiouridine, were tested to investigate the selectivity of the MTSEA biotin-XX s4U detection system. The system did not react with 5-methyl-2-thiouridine in tRNAs from a T. thermophilus tRNA 4-thiouridine synthetase (thiI) gene deletion strain. Thus, the most useful advantage of the MTSEA biotin-XX s4U detection system is that MTSEA biotin-XX reacts only with s4U and not with other sulfur-containing modified nucleosides such as s2U derivatives in tRNAs. Furthermore, the MTSEA biotin-XX s4U detection system can analyze multiple samples in a short time span. The MTSEA biotin-XX s4U detection system can also be used for the analysis of s4U formation in tRNA. Finally, we demonstrate that the MTSEA biotin-XX system can be used to visualize newly transcribed tRNAs in S. cerevisiae cells.