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  • P-ISSN2233-4203
  • E-ISSN2093-8950
  • ESCI, SCOPUS, KCI

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  • P-ISSN 2233-4203
  • E-ISSN 2093-8950

Artificial Oxidation of Cysteine Residues in Peroxiredoxin 6 Detected by Twodimensional Gel Electrophoresis and Capillary Liquid Chromatography- Electrospray Mass Spectrometry

Mass Spectrometry Letters / Mass Spectrometry Letters, (P)2233-4203; (E)2093-8950
2012, v.3 no.1, pp.10-14
https://doi.org/10.5478/MSL.2012.3.1.010
Junko Kimata (Thermo Fisher Scientific)
Yasushi Shigeri (National Institute of Advanced Industria)
Yasukazu Yoshida (National Institute of Advanced Industria)
Etsuo Niki (National Institute of Advanced Industria)
Tomoya Kinumi (Advanced Industrial Science and Technolo)
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Abstract

Artificially oxidized cysteine residues in peroxiredoxin 6 (Prx6) were detected by electrospray interface capillary liquidchromatography-linear ion trap mass spectrometry after the preparation of two-dimensional gel electrophoresis (2D-GE). Weused Prx6 as a model protein because it possesses only two cysteine residues at the 47th and 91st positions. The spot of Prx6 on2D-GE undergoes a basic (isoelectric point, pI 6.6) to acidic (pI 6.2) shift by exposure to peroxide due to selective overoxidationof the active-site cysteine Cys-47 but not of Cys-91. However, we detected a tryptic peptide containing cysteine sulfonic acid atthe 47th position from the basic spot and a peptide containing both oxidized Cys-47 and oxidized Cys-91 from the acidic spot ofPrx6 after the separation by 2D-GE. We prepared two types of oxidized Prx6s: carrying oxidized Cys-47 (single oxidized Prx6),and other carrying both oxidized Cys-47 and Cys-91 (double oxidized Prx6). Using these oxidized Prx6s, the single oxidizedPrx6 and double oxidized Prx6 migrated to pIs at 6.2 and 5.9, respectively. These results suggest that oxidized Cys-47 from thebasic spot and oxidized Cys-91 from the acidic spot are generated by artificial oxidation during sample handling processes afterisoelectric focusing of 2D-GE. Therefore, it is important to make sure of the origin of cysteine oxidation, if it is physiological orartificial, when an oxidized cysteine residue(s) is identified.

keywords
Two-dimensional gel electrophoresis, Peroxiredoxin, Cysteine sulfonic acid, Capillary HPLC-MS/MS, MALDITOF MS


참고문헌

1

Lopez, J. L. (2007). . J. Chromatogr. B, 849, 190-.

2

Kinumi, T. (2005). . Free Radic. Res, 39, 1335-.

3

Sultana, R. (2006). . Neurobiol. Aging, 27, 1564-.

4

Yoshida, Y. (2008). . Neurobiol. Aging, 30, 174-.

5

Poole, L. B. (1989). . J. Biol. Chem, 264, 12330-.

6

Fu, X. (2001). . J. Biol. Chem, 276, 41279-.

7

Rabilloud, T. (2002). . J. Biol. Chem, 277, 19396-.

8

Kinumi, T. (2004). . Biochem. Biophys. Res. Commun, 317, 722-.

9

Jia, Y. (2007). . J. Biol. Chem, 282, 4894-.

10

Rhee, S. G. (2005). . Free Radic. Biol. Med, 38, 1543-.

11

Manevich, Y. (2004). . Proc. Natl. Acad. Sci. USA, 101, 3780-.

12

Wagner, E. (2002). . Biochem. J, 366, 777-.

13

Kinumi, T. (2006). . J. Mass Spectrom, 41, 103-.

14

Tojo, H. (2004). . J. Chromatogr. A, 1056, 223-.

15

Muller, D. R. (1999). . J. Mass Spectrom, 34, 336-.

16

Herbert, B. (2001). . Electrophoresis, 22, 2046-.

17

Hamdan, M. (2001). . Mass Spectrom. Rev, 20, 121-.

18

Plowman, J. E. (2003). . Proteomics, 3, 942-.

19

Chiari, M. (1991). . Electrophoresis, 12, 376-.

20

Sun, G. (2004). . Electrophoresis, 25, 959-.

투고일Submission Date
2011-12-12
수정일Revised Date
2012-02-21
게재확정일Accepted Date
2012-02-21
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