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Assay for methylmalonyl coenzyme A mutase activity based on determination of succinyl coenzyme A by ultrahigh-performance liquid chromatography tandem mass spectrometry

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Abstract

Methylmalonic acidemia (MMA) is an inherited metabolic disease. In this condition, metabolism from methylmalonyl coenzyme A (CoA) to succinyl-CoA is inhibited because of either low methylmalonyl-CoA mutase (MCM) activity or adenosylcobalamin deficiency owing to altered vitamin B12 metabolism. A high-precision assay for detecting MCM activity would facilitate not only MMA diagnosis but also the ability to determine the severity of MMA. We developed an MCM assay method based on ultrahigh-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) that involves the determination of succinyl-CoA, which is formed in an enzyme reaction, using peripheral lymphocytes. Using 0.05, 0.5, and 5 μmol/L succinyl-CoA, the intra-assay coefficient of variation (CV) was less than 5.2 % and the inter-assay CV was less than 8.7 %. The MCM activities of five healthy individuals and four patients were investigated with this assay. The MCM activities of the patients were very low in relation to those of healthy individuals. Together, these results show that the UPLC–MS/MS method is useful for a detailed MCM activity assay.

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References

  1. Fowler B, Leonard JV, Baumqartner MR (2008) Causes of and diagnostic approach to methylmalonic acidurias. J Inherit Metab Dis 31:350–360

    Article  CAS  Google Scholar 

  2. Manoli I, Venditti CP (2005) Methylmalonic acidemia. In: Pagon RA, Adam MP, Ardinger HH, Bird TD, Dolan CR, Fong CT, Smith RJH, Stephens K (eds) Gene reviews. University of Washington, Seattle, pp 1993–2015

    Google Scholar 

  3. Fenton WA, Gravel RA, Rosenblatt DS (2001) Disorders of propionate and methylmalonate metabolism. In: Charles S, Arthur B, William S, David V, Barton C, Kenneth K, Bert V (eds) The metabolic and molecular bases of inherited disease, 8th edn. McGraw-Hill, New York, pp 2165–2193

    Google Scholar 

  4. Fujisawa D, Nakamura K, Mitsubuchi H, Ohura T, Shigematsu Y, Yorifuji T, Kasahara M, Horikawa R, Endo F (2013) Clinical features and management of organic acidemias in Japan. J Hum Genet 58:769–774

    Article  Google Scholar 

  5. Cornejo V, Colombo M, Duran G, Mabe P, Jimenez M, De la Parra A, Raimann E (2002) Diagnosis and follow up of 23 children with organic acidurias. Rev Med Chil 130:259–266

    Google Scholar 

  6. Millington DS, Kodo N, Norwood DL, Roe CR (1990) Tandem mass spectrometry: a new method for acylcarnitine profiling with potential for neonatal screening for inborn errors of metabolism. J Inherit Metab Dis 13:321–324

    Article  CAS  Google Scholar 

  7. Rashed MS, Ozand PT, Bucknall MP, Little D (1995) Diagnosis of inborn errors of metabolism from blood spots by acylcarnitines and amino acids profiling using automated electrospray tandem mass spectrometry. Pediatr Res 38:324–331

    Article  CAS  Google Scholar 

  8. Dionisi-Vici C, Deodato F, Roschinger W, Rhead W, Wilcken B (2006) Classical organic acidurias, propionic aciduria, methylmalonic aciduria and isovaleric aciduria: long-term outcome and effects of expanded newborn screening using tandem mass spectrometry. J Inherit Metab Dis 29:383–389

    Article  CAS  Google Scholar 

  9. Shigemateu Y, Hirano S, Hata I, Tanaka Y, Sudo M, Sakura N, Tajima T, Yamaguchi S (2002) Newborn mass screening and selective screening using electrospray tandem mass spectrometry in Japan. J Chromatoqr B Anal Technol Biomed Life Sci 776:39–48

    Article  Google Scholar 

  10. Nakagawa K, Kawana S, Hasegawa Y, Yamaguchi S (2010) Simplified method for the chemical diagnosis of organic aciduria using GC/MS. J Chromatogr B 878:942–948

    Article  CAS  Google Scholar 

  11. Morrow G III, Mahoney MJ, Mathews C, Lebowitz J (1975) Studies of methylmalonyl coenzyme A carbonylmutase activity in methylmalonic acidemia. I. Correlation of clinical, hepatic, and fibroblast data. Pediatr Res 9:641–644

    Article  CAS  Google Scholar 

  12. Kikuchi M, Hanamizu H, Narisawa K, Tada K (1989) Assay of methylmalonyl CoA mutase with high-performance liquid chromatography. Clin Chim Acta 184:307–314

    Article  CAS  Google Scholar 

  13. Tajima G, Yofune H, Bahagia Febriani AD, Nishimura Y, Ono H, Sakura N (2004) A simple and rapid enzymatic assay for the branched-chain α-ketoacid dehydrogenase complex using high-performance liquid chromatography. J Inherit Metab Dis 27:633–639

    Article  CAS  Google Scholar 

  14. Tajima G, Sakura N, Yofune H, Nishimura Y, Ono H, Hasegawa Y, Hata I, Kimura M, Yamaguchi S, Shigematsu Y, Kobayashi M (2005) Enzymatic diagnosis of medium-chain acyl-CoA dehydrogenase deficiency by detecting 2-octenoyl-CoA production using high-performance liquid chromatography: a practical confirmatory test for tandem mass spectrometry newborn screening in Japan. J Chromatogr B 823:122–130

    Article  CAS  Google Scholar 

  15. Peters HL, Nefedov M, Lee LW, Abdenur JE, Chamoles NA, Kahler SG, Ioannou PA (2002) Molecular studies in mutase-deficient (MUT) methylmalonic aciduria: identification of five novel mutations. Hum Mutat 20:406

    Article  Google Scholar 

  16. Sakamoto O, Ohura T, Matsubara Y, Takayanagi M, Tsuchiya S (2007) Mutation and haplotype analyses of the MUT gene in Japanese patients with methylmalonic acidemia. J Hum Genet 52:48–55

    Article  CAS  Google Scholar 

  17. Liu MY, Liu TT, Yang YL, Chang YC, Fan YL, Lee SF, Teng YT, Chiang SH, Niu DM, Lin SJ, Chao MC, Lin SP, Han LS, Qi Y, Hsiao KJ (2012) Mutation profile of the MUT gene in Chinese methylmalonic aciduria patients. JIMD Rep 6:55–64

    Google Scholar 

  18. Ghoraba DA, Mohammed MM, Zaki OK (2015) Mutation analysis of methylmalonyl CoA mutase gene exon 2 in Egyptian families: identification of 25 novel allelic variants. Meta Gene 3:71–88

    Article  Google Scholar 

  19. Gao L, Chiou W, Tang H, Cheng X, Camp HS, Burns DJ (2007) Simultaneous quantification of malonyl-CoA and several other short-chain acyl-CoAs in animal tissues by ion-pairing reversed-phase HPLC/MS. J Chromatogr B 853:303–313

    Article  CAS  Google Scholar 

  20. Dean JA (1992) Lange’s handbook of chemistry, 14th edn. McGraw-Hill, New York

    Google Scholar 

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Acknowledgments

This research was supported by a Grant-in-Aid for Research in Nagoya City University.

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Authors and Affiliations

  1. Department of Hospital Pharmacy, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya, 467-8603, Japan

    Kana Gotoh, Yuji Hotta, Yoshihiro Kawade, Kazunori Kimura & Yasuhiro Maeda

  2. Department of Pediatrics, School of Medicine, Fujita Health University, 1-98 Dengakugakubo, Kutsukake-cho, Toyoake, 470-1192, Japan

    Yoko Nakajima & Tetsuya Ito

  3. Department of Pediatrics, Hiroshima University Graduate School of Biomedical and Health Sciences, 1-2-3 Kasumi, Minami-ku, Hiroshima, 734-8551, Japan

    Go Tajima

  4. Department of Clinical Pharmaceutics, Graduate School of Medical Sciences, Nagoya City University, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya, 467-8601, Japan

    Tomoya Kataoka & Kazunori Kimura

  5. Department of Pediatrics, School of Pharmacy, Aichi-Gakuin University, 1-100 Kusumoto-cho, Chikusa-ku, Nagoya, 464-8650, Japan

    Naruji Sugiyama

Authors
  1. Kana Gotoh
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  2. Yoko Nakajima
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  3. Go Tajima
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  4. Yuji Hotta
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  5. Tomoya Kataoka
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  6. Yoshihiro Kawade
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  7. Naruji Sugiyama
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  8. Tetsuya Ito
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  9. Kazunori Kimura
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  10. Yasuhiro Maeda
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Corresponding author

Correspondence to Yasuhiro Maeda.

Additional information

Published in the topical collection New Applications of Mass Spectrometry in Biomedicine with guest editors Fumio Nomura, Mitsutoshi Setou, and Toshimitsu Niwa.

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Gotoh, K., Nakajima, Y., Tajima, G. et al. Assay for methylmalonyl coenzyme A mutase activity based on determination of succinyl coenzyme A by ultrahigh-performance liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 407, 5281–5286 (2015). https://doi.org/10.1007/s00216-015-8753-8

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  • DOI: https://doi.org/10.1007/s00216-015-8753-8

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