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Complete amino acid sequencing and immunoaffinity clean-up can facilitate screening of various chemical modifications on human serum albumin

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Abstract

This manuscript describes a simple and practical strategy for screening of various chemical modifications of human serum albumin (HSA). Serum albumin is the most abundant blood plasma protein in humans (HSA, 66.5 kDa, t1/2 = 19 d), constituting about 60 % of total proteins. Therefore, it is believed to be the main target of chemical stresses during physiological events such as increased oxidative stress from the degenerative diseases of aging, and higher glucose stress in diabetes mellitus. Consequently, chemical modifications can provide significant information about these biological events. In this study, a complete and robust sequencing method was attained by the peptide mass fingerprinting (PMF) technique using two different complementary proteases (trypsin and Glu-C) and matrix-assisted laser desorption ionization-time of flight/mass spectrometry (MALDI-TOF/MS) in both positive and negative ionization modes. Using this strategy, several modified peptides, 12 oxidations, 25 glycations, 6 lipoxidations, and 5 nitrations have been identified on HSA treated with chemical reactions in vitro. Combined with immunoaffinity clean-up, this method was able to detect in vivo chemical modifications of HSA and found oxidized Trp214 and glycated Lys525 in healthy human plasma.

Complete amino acid sequencing can facilitate screening of chemical modifications on HSA

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Abbreviations

2D:

two-dimensional

ACTH:

adrenocorticotropic hormone

Ang:

angiotensin

DHB:

2,5-dihydroxybenzoic acid

DTT:

dithiothreitol

ELISA:

enzyme-linked immunosorbent assay

ESI:

electrospray ionization

HNE:

4-hydroxy-2(E)-nonenal

HSA:

human serum albumin

IAA:

iodoacetamide

Ins B:

insulin chain B oxidized, bovine

LC:

liquid chromatography

MALDI:

matrix-assisted laser desorption ionization

MS:

mass spectrometry

PMF:

peptide mass fingerprinting

RNS:

reactive nitrogen species

ROS:

reactive oxygen species

TFA:

trifluoroacetic acid

TOF:

time-of-flight

V8:

endoproteinase Glu-C

References

  1. He QY, Chiu JF (2003) Proteomics in biomarker discovery and drug development. J Cell Biochem 89:868–886

    Article  CAS  Google Scholar 

  2. Anderson NL, Anderson NG (2002) The human plasma proteome: history, character, and diagnostic prospects. Mol Cell Proteomics 1:845–867

    Article  CAS  Google Scholar 

  3. Tu C, Rudnick PA, Martinez MY, Cheek KL, Stein SE et al (2010) Depletion of abundant plasma proteins and limitations of plasma proteomics. J Proteome Res 9:4982–4991

    Article  CAS  Google Scholar 

  4. Peters T Jr (1996) All About Albumin: Biochemistry, Genetics, and Medical Applications. Academic Press, San Diego, CA. ISBN 0-12-552110-3

    Google Scholar 

  5. Goto T, Kojima S, Shitamichi S, Lee SH, Oe T (2012) Chemical modificomics: a novel strategy for efficient biomarker discovery through chemical modifications on a target peptide. Anal Methods 4:1945–1952

    Article  CAS  Google Scholar 

  6. Lee SH, Goto T, Oe T (2008) A novel 4-oxo-2(E)-nonenal-derived modification to angiotensin II: oxidative decarboxylation of N-terminal aspartic acid. Chem Res Toxicol 21:2237–2244

    Article  CAS  Google Scholar 

  7. Lee SH, Takahashi R, Goto T, Oe T (2010) Mass spectrometric characterization of modifications to angiotensin II by lipid peroxidation products, 4-oxo-2(E)-nonenal and 4-hydroxy-2(E)-nonenal. Chem Res Toxicol 23:1771–1785

    Article  CAS  Google Scholar 

  8. Lee SH, Masuda T, Goto T, Oe T (2013) MALDI-TOF/MS-based label-free binding assay for angiotensin II type 1 receptor: application for novel angiotensin peptides. Anal Biochem 437:10–16

    Article  CAS  Google Scholar 

  9. Inoue K, Garner C, Ackermann BL, Oe T, Blair IA (2006) Liquid chromatography/tandem mass spectrometry characterization of oxidized amyloid beta peptides as potential biomarkers of Alzheimer’s disease. Rapid Commun Mass Spectrom 20:911–918

    Article  CAS  Google Scholar 

  10. Trostchansky A, Lind S, Hodara R, Oe T, Blair IA, Ischiropoulos H et al (2006) Interaction with phospholipids modulates alpha-synuclein nitration and lipid-protein adduct formation. Biochem J 393:343–349

    Article  CAS  Google Scholar 

  11. Silva CJ, Onisko BC, Dynin I, Erickson ML, Vensel WH, Requena JR et al (2010) Assessing the role of oxidized methionine at position 213 in the formation of prions in hamsters. Biochemistry 49:1854–1861

    Article  CAS  Google Scholar 

  12. Lee SH, Miyamoto K, Goto T, Oe T (2011) Noninvasive proteomic analysis of human skin keratins: screening of methionine oxidation in keratins by mass spectrometry. J Proteomics 75:435–449

    Article  CAS  Google Scholar 

  13. Otagiri M, Chuang VT (2009) Pharmaceutically important pre- and post-translational modifications on human serum albumin. Biol Pharm Bull 32:527–534

    Article  CAS  Google Scholar 

  14. Alex Merrick B (2008) The plasma proteome, adductome, and idiosyncratic toxicity in toxicoproteomics research. Brief Funct Genomic Proteomic 7:35–49

    Article  Google Scholar 

  15. Rubino FM, Pitton M, Di Fabio D, Colombi A (2009) Toward an “omic” physiopathology of reactive chemicals: thirty years of mass spectrometric study of the protein adducts with endogenous and xenobiotic compounds. Mass Spectrom Rev 28:725–784

    Article  CAS  Google Scholar 

  16. Roohk HV, Zaidi AR (2008) A review of glycated albumin as an intermediate glycation index for controlling diabetes. J Diabetes Sci Technol 2:1114–1121

    Google Scholar 

  17. Aldini G, Regazzoni L, Orioli M, Rimoldi I, Facino RM, Carini M (2008) A tandem MS precursor-ion scan approach to identify variable covalent modification of albumin Cys34: A new tool for studying vascular carbonylation. J Mass Spectrom 43:1470–1481

    Article  CAS  Google Scholar 

  18. Sanaki T, Suzuki M, Lee SH, Goto T, Oe T (2010) A simple and efficient approach to improve protein identification by the peptide mass fingerprinting method: concomitant use of negative ionization. Anal Methods 2:1144–1151

    Article  CAS  Google Scholar 

  19. Finch JW, Crouch RK, Knapp DR, Schey KL (1993) Mass spectrometric identification of modifications to human serum albumin treated with hydrogen peroxide. Arch Biochem Biophys 305:595–599

    Article  CAS  Google Scholar 

  20. Marx G, Chevion M (1986) Site-specific modification of albumin by free radicals. Reaction with copper(II) and ascorbate. Biochem J 236:397–400

    CAS  Google Scholar 

  21. Gadgil HS, Bondarenko PV, Treuheit MJ, Ren D (2007) Screening and sequencing of glycated proteins by neutral loss scan LC/MS/MS method. Anal Chem 79:5991–5999

    Article  CAS  Google Scholar 

  22. Aldini G, Gamberoni L, Orioli M, Beretta G, Regazzoni L, Maffei Facino R et al (2006) Mass spectrometric characterization of covalent modification of human serum albumin by 4-hydroxy-trans-2-nonenal. J Mass Spectrom 41:1149–1161

    Article  CAS  Google Scholar 

  23. Jiao K, Mandapati S, Skipper PL, Tannenbaum SR, Wishnok JS (2001) Site-selective nitration of tyrosine in human serum albumin by peroxynitrite. Anal Biochem 293:43–52

    Article  CAS  Google Scholar 

  24. Wa C, Cerny R, Hage DS (2006) Obtaining high sequence coverage in matrix-assisted laser desorption time-of-flight mass spectrometry for studies of protein modification: analysis of human serum albumin as a model. Anal Biochem 349:229–241

    Article  CAS  Google Scholar 

  25. Lee SH, Oe T, Blair IA (2001) Vitamin C-induced decomposition of lipid hydroperoxides to endogenous genotoxins. Science 292:2083–2086

    Article  CAS  Google Scholar 

  26. Danielson SR, Andersen JK (2008) Reactive nitrogen species. Oxidative and nitrative protein modifications in Parkinson’s disease. Free Radical Biol Med 44:1787–1794

    Article  CAS  Google Scholar 

  27. Shen Y, Jacobs JM, Camp DG II, Fang R, Moore RJ, Smith RD et al (2004) Ultra-high-efficiency strong cation exchange LC/RPLC/MS/MS for high dynamic range characterization of the human plasma proteome. Anal Chem 76:1134–1144

    Article  CAS  Google Scholar 

  28. Oe T, Arora JS, Lee SH, Blair IA (2003) A novel lipid hydroperoxide-derived cyclic covalent modification to histone H4. J Biol Chem 278:42098–42105

    Article  CAS  Google Scholar 

  29. Yocum AK, Oe T, Yergey AL, Blair IA (2005) Novel lipid hydroperoxide-derived hemoglobin histidine adducts as biomarkers of oxidative stress. J Mass Spectrom 40:754–764

    Article  CAS  Google Scholar 

  30. Geoghegan KF, Hoth LR, Tan DH, Borzilleri KA, Withka JM, Boyd JG (2002) Cyclization of N-terminal S-carbamoylmethylcysteine causing loss of 17 Da from peptides and extra peaks in peptide maps. J Proteome Res 1:181–187

    Article  CAS  Google Scholar 

  31. Finley EL, Dillon J, Crouch RK, Schey KL (1998) Identification of tryptophan oxidation products in bovine alpha-crystallin. Protein Sci 7:2391–2397

    Article  CAS  Google Scholar 

  32. Patterson LK, Mazière JC, Bartels DM, Hug GL, Santus R, Morlière P (2012) Evidence for a slow and oxygen-insensitive intra-molecular long range electron transfer from tyrosine residues to the semioxidized tryptophan 214 in human serum albumin: its inhibition by bound copper (II). Amino Acids 42:1269–1275

    Article  CAS  Google Scholar 

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Acknowledgments

This work was supported in part by Grants-in-Aid for Challenging Exploratory Research (to T.O., # 21659035 and # 23659016), a Grant-in-Aid for Young Scientists (B) (to T.G., # 22790032), a Grant-in-Aid for Scientific Research (C) (to T.G., # 40344684) from Japan Society for the Promotion of Science (JSPS), and a research grant (to S.H.L. for 2009) from the Suzuken Memorial Foundation (Nagoya, Japan). The authors thank the Biomedical Research Core, School of Medicine in Tohoku University for the use of MALDI-TOF/MS.

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Authors and Affiliations

  1. Department of Bioanalytical Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, Aobayama, Aoba-ku, Sendai, 980-8578, Japan

    Takaaki Goto, Kazuyuki Murata, Seon Hwa Lee & Tomoyuki Oe

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  1. Takaaki Goto
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  2. Kazuyuki Murata
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  3. Seon Hwa Lee
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  4. Tomoyuki Oe
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Correspondence to Tomoyuki Oe.

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Goto, T., Murata, K., Lee, S.H. et al. Complete amino acid sequencing and immunoaffinity clean-up can facilitate screening of various chemical modifications on human serum albumin. Anal Bioanal Chem 405, 7383–7395 (2013). https://doi.org/10.1007/s00216-013-7146-0

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  • DOI: https://doi.org/10.1007/s00216-013-7146-0

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