Abstract
Aim:
To study the non-covalent interaction between glutathione and common amino acids.
Methods:
A stoichiometry of glutathione and common amino acids were mixed to reach the equilibrium, and then the mixed solution was investigated by electrospray ionization mass spectrometry (ESI–MS). The binding of the complexes was further examined by collision-induced dissociation (CID) in a tandem mass spectrometer as well as UV spectroscopy. To avoid distinct ionization efficiency discrepancy and signal suppression in the ESI-MS measurements, the interaction between glutathione (GSH) and glutamate (Glu) was quantitatively evaluated. The total concentrations and series of m/z of peak intensities for glutathione and amino acids could be achieved, respectively. Due to the existence of some oligomeric species arising from glutathione or amino acids, an improved calculation formula was proposed to calculate the dissociation constants of glutathione binding to amino acids.
Results:
The ESI mass spectra revealed that glutathione could interact easily with Met, Phe, Tyr, Ser, or Ile to form non-covalent complexes. The binding of the complexes was further confirmed by CID experiments in a tandem mass spectrometer as well as UV spectroscopy. Moreover, an improved calculation formula was successfully applied to determine the dissociation constants of glutathione binding to Glu, His, or Gln. Finally, a possible formation mechanism for the complexes of glutathione with amino acids was proposed.
Conclusion:
The reduced polypeptide γ-glutathione can interact with each of 8 common amino acids, including Glu, His, and Gln to form non-covalent complexes with different affinity.
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Konig S, Hasche A, Pallast S, Krieglstein J, Klumpp S . Detection of ATP-binding to growth factors. J Am Soc Mass Spectrom 2008; 19: 91–5.
Yanes O, Aviles FX, Roepstorff P, Nazabal A, Zenobi R, Calvete JJ . Exploring the “intensity fading” phenomenon in the study of non-covalent interactions by MALDI-TOF mass spectrometry. J Am Soc Mass Spectrom 2007; 18: 359–67.
Fabris D, Chaudhari P, Hagan N, Turner K . Functional investigations of retroviral protein-ribonucleic acid complexes by nanospray Fourier transform ion cyclotron resonance mass spectrometry. Eur J Mass Spectrom 2007; 13: 29–33.
Wilson JJ, Brodbelt JS . Infrared multiphoton dissociation of duplex DNA/drug complexes in a quadrupole ion trap. Anal Chem 2007; 79: 2067–77.
Baytekin B, Baytekin HT, Schalley CA . Mass spectrometric studies of non-covalent compounds: why supramolecular chemistry in the gas phase? Organ Biomol Chem 2006; 4: 2825–41.
Bovet C, Wortmann A, Eiler S, Granger F, Ruff M, Gerrits B, et al. Estrogen receptor-ligand complexes measured by chip-based nanoelectrospray mass spectrometry: An approach for the screening of endocrine disruptors. Protein Sci 2007; 16: 938–46.
Lin ZPJ, Li WK, Dai GW . Application of LC-MS for quantitative analysis and metabolite identification of therapeutic oligonucleotides. J Pharmaceut Biomed Anal 2007; 44: 330–41.
Li MH, Yin LL, Cai MJ, Zhang WY, Huang Y, Wang X, et al. Design, synthesis, and anti-inflammatory evaluation of a series of novel amino acid-binding 1,5-diarylpvrazole derivatives. Acta Pharmacol Sin 2005; 26: 865–72.
Terrier P, Tortajada J, Buchmann W . A study of non-covalent complexes involving single-stranded DNA and polybasic compounds using nanospray mass spectrometry. J Am Soc Mass Spectrom 2007; 18: 346–58.
Mazzitelli CL, Brodbelt JS . Probing ligand binding to duplex DNA using KMnO4 reactions and electrospray ionization tandem mass spectrometry. Anal Chem 2007; 79: 4636–47.
Lu HJ, Guo YL, Yang PY . Using amino acids for probing structure information of cytochrome c by electrospray ionization mass spectrometry. J Am Soc Mass Spectrom 2004; 15: 1612–5.
Lear BJ, Kubiak CP . Origins of cooperative non-covalent host-guest chemistry in mixed valence complexes. J Phys Chem B 2007; 111: 6766–71.
Li Y, Zhong DF, Chen SW, Maeba I . Identification of some benproperine metabolites in humans and investigation of their antitussive effect. Acta Pharmacol Sin 2005; 26: 1519–26.
Bligh SWA, Haley T, Lowe PN . Measurement of dissociation constants of inhibition binding to Src-2 SH2 domain protein by non-covalent electrospray ionization mass spectrometry. J Mol Recognit 2003; 16: 139–47.
Yu Z, Cui M, Yan CY, Song FR, Liu ZQ, Liu SY . Investigation of heptakis (2,6-di-O-methyl)-beta-cyclodextrin inclusion complexes with flavonoid glycosides by electrospray ionization mass spectrometry. Rapid Commun Mass Spectrom 2007; 21: 683–90.
Wang W, Kitova EN, Klassen JS . Bioactive recognition sites may not be energetically preferred in protein-carbohydrate complexes in the gas phase. J Am Chem Soc 2003; 125: 13 630–1.
Wigger M, Eyler JR, Benner SA, Li WQ, Marshall AG . Fourier transform-ion cyclotron resonance mass spectrometric resolution, identification, and screening of non-covalent complexes of Hck Src homology 2 domain receptor and ligands from a 324-member peptide combinatorial library. J Am Soc Mass Spectrom 2002; 13: 1162–9.
Nyadong L, Green MD, De Jesus VR, Newton PN, Fernandez FM . Reactive desorption electrospray ionization linear ion trap mass spectrometry of latest-generation counterfeit antimalarials via non-covalent complex formation. Anal Chem 2007; 79: 2150–7.
Baydas G, Sonkaya E, Tuzcu M, Yasar A, Donder E . Novel role for gabapentin in neuroprotection of central nervous system in streptozotocine-induced diabetic rats. Acta Pharmacol Sin 2005; 26: 417–22.
Levtchenko E, De Graaf-Hess A, Wilmer M, Van den Heuvel L, Monnens L, Blom H . Altered status of glutathione and its metabolites in cystinotic cells. Nephrol Dial Transplant 2005; 20: 1828–32.
Sheng R, Gu ZL, Xie ML, Zhou WX, Guo CY . EGCG inhibits cardiomyocyte apoptosis in pressure overload-induced cardiac hypertrophy and protects cardiomyocytes from oxidative stress in rats. Acta Pharmacol Sin 2007; 28: 191–201.
Chin TY, Chueh SH, Tao PL . S-nitrosoglutathione and glutathione act as NMDA receptor agonists in cultured hippocampal neurons. Acta Pharmacol Sin 2006; 27: 853–60.
Njalssson R, Norgren S . Physiological and pathological aspects of GSH metabolism. Acta Paediat 2005; 94: 132–7.
Schmidt AC, Neustadt M, Otto M . Quantitative evaluation of the binding of phenylarsenic species to glutathione, isotocin, and thioredoxin by means of electrospray ionization time-of-flight mass spectrometry. J Mass Spectrom 2007; 42: 771–80.
Yao X, Wang YT, Chen G . Simultaneous determination of aminothiols, ascorbic acid and uric acid in biological samples by capillary electrophoresis with electrochemical detection. Biomed Chromatogr 2007; 21: 520–6.
Zhang HR, Chen G, Wang L, Ding L, Tian Y, Jin WJ, et al. Study on the inclusion complex of cyclodexin and sulphonated azodyes by electrospray ionization mass spectrometry. Int J Mass Spectrom 2006; 252: 1–10.
Han MJ, Meng XG, Lippincott L . Determination of configuration of arsenite-glutathione complexes using ECSTM. Toxicol Lett 2007; 175: 57–63.
Kuipers BJH, Gruppen H . Prediction of molar extinction coefficients of proteins and peptides using UV absorption of the constituent amino acids at 214 nm to enable quantitative reverse phase high-performance liquid chromatography-mass spectrometry analysis. J Agric Food Chem 2007; 55: 5445–51.
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Project supported by the National Natural Science Foundation of China (No 20473020).
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Dai, Zy., Chu, Yq., Wu, B. et al. Investigation of non-covalent complexes of glutathione with common amino acids by electrospray ionization mass spectrometry. Acta Pharmacol Sin 29, 759–771 (2008). https://doi.org/10.1111/j.1745-7254.2008.00791.x
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DOI: https://doi.org/10.1111/j.1745-7254.2008.00791.x