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Low Mass MS/MS Fragments of Protonated Amino Acids Used for Distinction of Their 13C- Isotopomers in Metabolic Studies

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

Glu, Gln, Pro, and Ala are the main amino acids involved in ammonia detoxification in mosquitoes. In order to develop a tandem mass spectrometry method (MS2) to monitor each carbon of the above isotopically-labeled 13C-amino acids for metabolic studies, the compositions and origins of atoms in fragments of the protonated amino acid should be first elucidated. Thus, various electrospray (ESI)-based MS2 tools were employed to study the fragmentation of these unlabeled and isotopically-labeled amino acids and better understand their dissociation pathways. A broad range of fragments, including previously-undescribed low m/z fragments was revealed. The formulae of the fragments (from m/z 130 down to m/z 27) were confirmed by their accurate masses. The structures and conformations of the larger fragments of Glu were also explored by ion mobility mass spectrometry (IM-MS) and gas-phase hydrogen/deuterium exchange (HDX) experiments. It was found that some low m/z fragments (m/z 27–30) are common to Glu, Gln, Pro, and Ala. The origins of carbons in these small fragments are discussed and additional collision induced dissociation (CID) MS2 fragmentation pathways are proposed for them. It was also found that small fragments (≤m/z 84) of protonated, methylated Glu, and methylated Gln are the same as those of the underivatized Glu and Gln. Taken together, the new approach of utilizing low m/z fragments can be applied to distinguish, identify, and quantify 13C-amino acids labeled at various positions, either in the backbone or side chain.

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Acknowledgments

The authors thank Yang Song for her contribution in the early stages of this work, Mowei Zhou for his help on G2 Q-TOF, and George Tsaprailis and Yelena Feinstein for access to the AB/SCIEX 3000 QqQ and AB/SCIEX 4000 QTRAP mass spectrometers at the Arizona Proteomics Consortium, The University of Arizona. The AB/SCIEX 4000 QTRAP mass spectrometer was provided by NIH/NCRR Grant 1S10RR022384-01. The authors are also grateful to Dr. Yayoi Hongo (RIKEN, Wako, Japan) and David R. Bush (The University of Arizona) for fruitful discussions, and Dr. Kanamatsu (Soka University, Tokyo, Japan) and Dr. Takatori (Meiji Pharmaceutical University, Tokyo, Japan) for kindly providing [4-13C] – Gln and [4-13C] – Glu, respectively, which are not commercially available. This work was financially supported by NIH/NIAID Grant R01AI088092 (to PYS).

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Author notes

  1. Xin Ma & Vicki H. Wysocki

    Present address: Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, 43210, USA

  2. Shai Dagan

    Present address: Israel Institute for Biological Research, Ness Ziona, 74100, Israel

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, The University of Arizona, Tucson, AZ, 85721, USA

    Xin Ma, Shai Dagan, Árpád Somogyi, Vicki H. Wysocki & Patricia Y. Scaraffia

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  1. Xin Ma
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  2. Shai Dagan
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  3. Árpád Somogyi
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  4. Vicki H. Wysocki
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  5. Patricia Y. Scaraffia
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Correspondence to Patricia Y. Scaraffia.

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Ma, X., Dagan, S., Somogyi, Á. et al. Low Mass MS/MS Fragments of Protonated Amino Acids Used for Distinction of Their 13C- Isotopomers in Metabolic Studies. J. Am. Soc. Mass Spectrom. 24, 622–631 (2013). https://doi.org/10.1007/s13361-012-0574-9

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  • DOI: https://doi.org/10.1007/s13361-012-0574-9

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