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β-Lactoglobulin detected in human milk forms noncovalent complexes with maltooligosaccharides as revealed by chip-nanoelectrospray high-resolution tandem mass spectrometry

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Abstract

Cow’s milk protein allergy in exclusively breastfed infants, the main cause of food intolerance during the first 6 months of life, is triggered by the mother’s diet. β-Lactoglobulin (BLG) present in cow’s milk is one of the most potent allergens for newborns. Since no prophylactic treatment is available, finding ligands capable of binding BLG and reducing its allergenicity is currently the focus of research. In this work, an innovative methodology encompassing microfluidics based on fully automated chip-nanoelectrospray ionization (nanoESI), coupled with high-resolution mass spectrometry (MS) on a quadrupole time-of-flight (QTOF MS) instrument was developed. This platform was employed for the assessment of the noncovalent interactions between maltohexaose (Glc6) and β-lactoglobulin extracted from human milk upon deliberate intake of cow’s milk. The experiments were carried out in (+) ESI mode, using ammonium acetate (pH 6.0) as the buffer and also in pure water. In both cases, the MS analysis revealed the formation of BLG–Glc6 complex, which was characterized by top-down fragmentation in tandem MS (MS/MS) using collision-induced dissociation (CID). Our findings have a significant biomedical impact, indicating that Glc6 binds BLG under conditions mimicking the in vivo environment and therefore might represent a ligand, able to reduce its allergenicity.

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Abbreviations

BLG:

β-Lactoglobulin

CID:

Collision-induced dissociation

ESI:

Electrospray ionization

Glc6 :

Maltohexaose

HPLC:

High-performance liquid chromatography

MS:

Mass spectrometry/spectrometer

MS/MS:

Tandem mass spectrometry

Mr:

Molecular mass

QTOF MS:

Quadrupole time-of-flight mass spectrometry/spectrometer

References

  • Boyce JA, Assa’ad A, Burks AW, Jones SM, Sampson HA, Wood RA, Plaut M, Cooper SF, Fenton MJ, Arshad SH, Bahna SL, Beck LA, Byrd-Bredbenner C, Camargo CA Jr, Eichenfield L, Furuta GT, Hanifin JM, Jones C, Kraft M, Levy BD, Lieberman P, Luccioli S, McCall KM, Schneider LC, Simon RA, Simons FE, Teach SJ, Yawn BP, Schwaninger JM (2010) Guidelines for the diagnosis and management of food allergy in the United States: report of the NIAID-sponsored expert panel. J Allergy Clin Immunol 126:S1–S58

    Article  PubMed Central  PubMed  Google Scholar 

  • Bu G, Luo Y, Jing L, Zhang Y (2010) Reduced antigenicity of β-lactoglobulin by conjugation with glucose through controlled Maillard reaction conditions. Food Agric Immunol 21:143–156

    Article  CAS  Google Scholar 

  • Cederkvist FH, Zamfir AD, Bahrke S, Eijsink VG, Sørlie M, Peter-Katalinić J, Peter MG (2006) Identification of a high-affinity-binding oligosaccharide by (+) nanoelectrospray quadrupole time-of-flight tandem of a noncovalent enzyme-ligand complex. Angew Chem Int Ed 45:2429–2434

    Article  CAS  Google Scholar 

  • Cohen SL, Chait BT (1997) Mass spectrometry of whole proteins eluted from sodium dodecyl sulfate—polyacrylamide gel electrophoresis gels. Anal Biochem 247:257–267

    Article  CAS  PubMed  Google Scholar 

  • Ding X, Yang Y, Zhao S, Li Y, Wang Z (2011) Analysis of α-lactalbumin, β-lactoglobulin A and B in whey protein powder, colostrum, raw milk, and infant formula by CE and LC. Dairy Sci Technol 91:213–225

    Article  CAS  Google Scholar 

  • Domon B, Costello CE (1988) A systematic nomenclature for carbohydrate fragmentations in FAB-MS/MS spectra of glycoconjugates. Glycoconj J 5:397–409

    Article  CAS  Google Scholar 

  • Flangea C, Schiopu C, Capitan F, Mosoarca C, Manea M, Sisu E, Zamfir AD (2013) Fully automated chip-based nanoelectrospray combined with electron transfer dissociation for high throughput top-down proteomics. Central Eur J Chem 11:25–34

    Article  CAS  Google Scholar 

  • Giuffrida F, Elmelegy IM, Thakkar SK, Marmet C, Destaillats F (2014) Longitudinal evolution of the concentration of gangliosides GM3 and GD3 in human milk. Lipids 49:997–1004

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Gregory KE, Walker WA (2013) Immunologic factors in human milk and disease prevention in the preterm infant. Curr Pediatr Rep 1:222–228

    Article  Google Scholar 

  • Lönnerdal B (2013) Bioactive proteins in breast milk. J Paediatr Child Health 49:1–7

    Article  PubMed  Google Scholar 

  • Marincola FC, Dessì A, Corbu S, Reali A, Fanos V (2015) Clinical impact of human breast milk metabolomics. Clin Chim Acta pii S0009–8981(15):00085–00086

    Google Scholar 

  • Neyestani TR, Djalali M, Pezeshki M (2003) Isolation of alpha-lactalbumin, beta-lactoglobulin, and bovine serum albumin from cow’s milk using gel filtration and anion-exchange chromatography including evaluation of their antigenicity. Protein Expr Purif 29(2):202–208

    Article  CAS  PubMed  Google Scholar 

  • Ohtomo H, Konuma T, Utsunoiya H, Tsuge H, Ikeguchi M (2011) Structure and stability of Gyuba, a β-lactoglobulin chimera. Protein Sci 20(11):1867–1875

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Rachagani S, Gupta ID, Gupta N, Gupta SC (2006) Genotyping of ß-Lactoglobulin gene by PCR-RFLP in Sahiwal and Tharparkar cattle breeds. BMC Genet 7:31

    Article  PubMed Central  PubMed  Google Scholar 

  • Sarbu M, Ghiulai RM, Zamfir AD (2014) Recent developments and applications of electron transfer dissociation mass spectrometry in proteomics. Amino Acids 46:1625–1634

    Article  CAS  PubMed  Google Scholar 

  • Sawyer L, Kontopidis G (2000) The core lipocain, bovine beta-lactoglobulin. Biochim Biophys Acta 1482(1–2):136–148

    Article  CAS  PubMed  Google Scholar 

  • Schiopu C, Vukelić Ž, Capitan F, Kalanj-Bognar S, Sisu E, Zamfir AD (2012) Chip-nanoelectrospray quadrupole time-of-flight tandem mass spectrometry of meningioma gangliosides: a preliminary study. Electrophoresis 33:1778–1786

    Article  CAS  PubMed  Google Scholar 

  • Schneider SS, Aslebagh R, Wetie AGN, Sturgeon SR, Darie CC, Arcaro KF (2014) Using breast milk to assess breast cancer risk: the role of mass spectrometry-based proteomics. Adv Exp Med Biol 806:399–408

    Article  CAS  PubMed  Google Scholar 

  • Sisu E, Flangea C, Serb A, Zamfir AD (2011) Modern developments in mass spectrometry of chondroitin and dermatan sulfate glycosaminoglycans. Amino Acids 41:235–256

    Article  CAS  PubMed  Google Scholar 

  • Underwood MA, Kalanetra KM, Bokulich NA, Mirmiran M, Barile D, Tancredi DJ, German JB, Lebrilla CB, Mills DA (2014) Prebiotic oligosaccharides in premature infants. J Pediatr Gastroenterol Nutr 58:352–360

    Article  CAS  PubMed  Google Scholar 

  • Walker A (2010) Breast milk as the gold standard for protective nutrients. J Pediatr 156:S3–S7

    Article  CAS  PubMed  Google Scholar 

  • Yoshida T, Sasahara Y, Miyakawa S, Hattori M (2005) Reduced T cell response to β-Lactoglobulin by conjugation with acidic oligosaccharides. J Agric Food Chem 53:6851–6857

    Article  CAS  PubMed  Google Scholar 

  • Zamfir AD, Fabris D, Capitan F, Munteanu C, Vukelić Ž, Flangea C (2013) Profiling and sequence analysis of gangliosides in human astrocytoma by high-resolution mass spectrometry. Anal Bioanal Chem 405:7321–7335

    Article  CAS  PubMed  Google Scholar 

  • Ziegler J, Abel S (2014) Analysis of amino acids by HPLC/electrospray negative ion tandem mass spectrometry using 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) derivatization. Amino Acids 46:2799–2808

    Article  CAS  PubMed  Google Scholar 

  • Zsila F, Bikadi Z, Simonyi M (2002) Retinoic acid binding properties of the lipocain member β-Lactoglobulin studied by circular dichroism, electronic absorption spectroscopy and molecular modeling methods. Biochem Pharmacol 64:1651–1660

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by EU FP7 MARIE CURIE-PIRSES-GA-2010 269256 project, by the Romanian National Authority for Scientific Research through projects PN-II-ID-PCE-2011-3-0047, PN-II-PCCA-2011-142, PN-II-PCCA-2014-191 to A.D.Z and by “Victor Babes” University of Medicine and Pharmacy from Timisoara, Romania, through the Internal Grant 15250/19.12.2012 to F. C.

Author information

Authors and Affiliations

  1. Department of Obstetrics, Gynecology and Neonatology, Victor Babes University of Medicine and Pharmacy, Eftimie Murgu Square 2, 300041, Timisoara, Romania

    Florina Capitan & Constantin Ilie

  2. Faculty of Physics, West University of Timisoara, Vasile Parvan Blvd. 4, 300223, Timisoara, Romania

    Adrian C. Robu

  3. Mass Spectrometry Laboratory, National Institute for Research and Development in Electrochemistry and Condensed Matter, Plautius Andronescu Str. 1, 300224, Timisoara, Romania

    Adrian C. Robu & Alina D. Zamfir

  4. Department of Paediatrics, Victor Babes University of Medicine and Pharmacy, Eftimie Murgu Square 2, 300041, Timisoara, Romania

    Catalin Schiopu

  5. Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, 1230 York Avenue, New York, NY, 10065-6399, USA

    Brian T. Chait

  6. Department of Chemistry, Laboratory of Analytical Chemistry and Biopolymer Structure Analysis, University of Konstanz, Universitaetsstrasse 10, 78457, Constance, Germany

    Michael Przybylski

  7. Laboratory for the Analysis and Modeling of Biological Systems, Aurel Vlaicu University of Arad, Revolutiei Blvd. 77, 310130, Arad, Romania

    Alina D. Zamfir

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  1. Florina Capitan
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  2. Adrian C. Robu
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  3. Catalin Schiopu
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  4. Constantin Ilie
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  5. Brian T. Chait
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  6. Michael Przybylski
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  7. Alina D. Zamfir
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Corresponding author

Correspondence to Alina D. Zamfir.

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The authors declare no conflicts of interest.

Ethical standards

This study has been approved by the Ethics Commission of Victor Babes University of Medicine and Pharmacy, Timisoara. Informed consent was obtained from all individual participants included in this research. All procedures on the human participants were in agreement with the ethical standards of Victor Babes University of Medicine and Pharmacy, Timisoara, National Authority for Scientific Research (ANCS, Romania) and with the 1964 Helsinki declaration and its later amendments.

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Handling Editor: P. R. Jungblut.

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Capitan, F., Robu, A.C., Schiopu, C. et al. β-Lactoglobulin detected in human milk forms noncovalent complexes with maltooligosaccharides as revealed by chip-nanoelectrospray high-resolution tandem mass spectrometry. Amino Acids 47, 2399–2407 (2015). https://doi.org/10.1007/s00726-015-2030-1

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  • DOI: https://doi.org/10.1007/s00726-015-2030-1

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