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Investigation of the structure of alpha-lactalbumin protein nanotubes using optical spectroscopy

Published online by Cambridge University Press:  19 December 2013

Özgür Tarhan ,
Enver Tarhan  and
Şebnem Harsa
Özgür Tarhan*
Affiliation:
Department of Food Engineering, Faculty of Engineering, Izmir Institute of Technology, Urla, Izmir TR-35430, Turkey
Enver Tarhan
Affiliation:
Department of Physics, Faculty of Science, Izmir Institute of Technology, Urla, Izmir TR-35430, Turkey
Şebnem Harsa
Affiliation:
Department of Food Engineering, Faculty of Engineering, Izmir Institute of Technology, Urla, Izmir TR-35430, Turkey
*
*For correspondence; e-mail: [email protected]
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Abstract

Alpha-lactalbumin (α-la) is one of the major proteins in whey. When partially hydrolysed with Bacillus licheniformis protease, it produces nanotubular structures in the presence of calcium ions by a self-assembly process. This study presents investigation of α-la protein structure during hydrolysis and nanotube formation using optical spectroscopy. Before spectroscopic measurements, nanotubes were examined with microscopy. The observed α-la nanotubes (α-LaNTs) were in the form of regular hollow strands with a diameter of about 20 nm and the average length of 1 μm. Amide and backbone vibration bands of the Raman spectra displayed remarkable conformational changes in α and β domains in the protein structure during nanotube growth. This was confirmed by the Fourier-transform infrared (FTIR) spectroscopy data. Also, FTIR analysis revealed certain bands at calcium (Ca++) binding sites of COO− groups in hydrolysed protein. These sites might be critical in nanotube elongation.

Keywords

α-Lactalbuminprotein nanotubesmicroscopyRamanFTIR
Type
Research Article
Information
Journal of Dairy Research , Volume 81 , Issue 1 , February 2014 , pp. 98 - 106
Copyright
Copyright © Proprietors of Journal of Dairy Research 2013 

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References

Ambrose, EJ & Elliot, A 1951 Infrared spectroscopic studies of globular protein structure. Proceedings of the Royal Society London Series A 208 75–90 Google Scholar
Anderle, G & Mendelsohn, R 1987 Thermal denaturation of globular proteins Fourier transform-infrared studies of the amide III spectral region. Biophysical Journal 52 69–74 CrossRefGoogle ScholarPubMed
Ashton, L & Blanch, EW 2010 pH-induced conformational transitions in α-lactalbumin investigated with two-dimensional Raman correlation variance plots and moving windows. Journal of Molecular Structure 947 132–138 CrossRefGoogle Scholar
Barron, LD, Blanch, EW & Hecht, L 2002 Unfolded proteins studied by Raman optical activity. Advanced in Protein Chemistry 62 51–90 CrossRefGoogle ScholarPubMed
Barth, A 2007 Infrared spectroscopy of proteins. Biochimica et Biophysica Acta 1767 1073–1101 CrossRefGoogle ScholarPubMed
Blanch, EW, Morozova-Roche, LA, Hecht, L, Noppe, W & Barron, LD 2000 Raman optical activity characterization of native and molten globule states of equine lysozyme: comparison with hen lysozyme and bovine α-lactalbumin. Biopolymers (Biospectroscopy) 57 235–248 3.0.CO;2-H>CrossRefGoogle ScholarPubMed
Breddam, K & Meldal, M 1992 Substrate preferences of glutamic-acid-specific endopeptidases assessed by synthetic peptide substrates based on intramolecular fluorescence quenching. European Journal of Blochemistry 206 103–107 CrossRefGoogle ScholarPubMed
Graveland-Bikker, JF & de Kruif, CG 2006 Unique milk protein based nanotubes: food and nanotechnology meet. Trends in Food Science and Technology 17 196–203 CrossRefGoogle Scholar
Graveland-Bikker, JF, Ipsen, R, Otte, J & de Kruif, CG 2004 Influence of calcium on the self-assembly of partially hydrolyzed α-lactalbumin. Langmuir 20 6841–6846 CrossRefGoogle ScholarPubMed
Graveland-Bikker, JF, Schaap, IAT, Schmidt, CF & de Kruif, CG 2006 Structural and mechanical study of a self-assembling protein nanotube. Nano Letters 6 616–621 CrossRefGoogle ScholarPubMed
Graveland-Bikker, JF, Koning, RI, Koerten, HK, Geels, RBJ, Heeren, RMA & de Kruif, CG 2009 Structural characterization of α-lactalbumin nanotubes. Soft Matter 5 2020–2026 CrossRefGoogle Scholar
Ikeda, S & Li-Chan, ECY 2004 Raman spectroscopy of heat-induced fine-stranded and particulate β-lactoglobulin gels. Food Hydrocolloids 18 489–498 CrossRefGoogle Scholar
Ipsen, R & Otte, J 2003 Nano-structuring by means of proteolysis rheology of novel gels from α-lactalbumin. Annual Transactions of the Nordic Rheology Society 11 89–93 Google Scholar
Ipsen, R & Otte, J 2007 Self-assembly of partially hydrolysed α-lactalbumin. Biotechnology Advances 25 602–605 CrossRefGoogle ScholarPubMed
Ipsen, R, Otte, J & Qvist, KB 2001 Molecular self-assembly of partially hydrolysed α-lactalbumin resulting in strong gels with a novel microstructure. Journal of Dairy Research 68 277–286 CrossRefGoogle ScholarPubMed
Ipsen, R, Otte, J & Qvist, KB 2003 Protease induced nano-tubular gels from α-lactalbumin. In Food Colloids, Biopolymers and Materials, Special Publication, Vol. 284, pp. 84–92 (Eds Dickinson, E & van Vliet, T). London: Royal Society of Chemistry Google Scholar
Jackson, M & Mantsch, HH 1995 The use and misuse of FTIR spectroscopy in the determination of protein structure. Critical Reviews in Biochemistry and Molecular Biology 30(2) 95–120 CrossRefGoogle ScholarPubMed
Kitagawa, T, Azuma, T & Hamaguchi, K 1979 The Raman spectra of Bence-Jones proteins. Disulfide stretching frequencies and dependence of Raman intensity of tryptophan residua on their environments. Biopolymers 18 451–465 CrossRefGoogle Scholar
Kong, J & Yu, S 2007 Fourier Transform Infrared Spectroscopic Analysis of protein secondary structures. Acta Biochimica et Biophysica Sinica 39 549–559 CrossRefGoogle ScholarPubMed
Krimm, S & Bandekar, J 1986 Vibrational spectroscopy and conformation of peptides, polypeptides, and proteins. Advanced in Protein Chemistry 38 181–367 CrossRefGoogle ScholarPubMed
Lord, RC & Yu, NT 1970 Laser-excited Raman spectroscopy of biomolecules. I. Native lysozyme and its constituent amino acids. Journal of Molecular Biology 50 509–524 CrossRefGoogle ScholarPubMed
Miura, T & Thomas, GJ Jr 1995 Raman spectroscopy of proteins and their assemblies. In Subcellular Biochemistry (Proteins: Structure, Function and Engineering), Vol. 24, pp. 55–99 (Eds Biswas, BB & Roy, S) New York: Plenum CrossRefGoogle Scholar
Miura, T, Takeuchi, H & Harada, I 1989 Tryptophan Raman bands sensitive to hydrogen bonding and side-chain conformation. Journal of Raman Spectroscopy 20 667–671 CrossRefGoogle Scholar
Mizuguchi, M, Nara, M, Kawano, K & Nitta, K 1997 FT-IR study of the Ca2+-binding to bovine α-lactalbumin. FEBS Letters 417 153–156 CrossRefGoogle ScholarPubMed
Nakanishi, M, Takesada, H & Tsuboi, M 1974 Conformation of the cystine linkages in bovine α-lactalbumin as revealed by its Raman effect. Journal of Molecular Biology 89 241–243 CrossRefGoogle ScholarPubMed
Nara, M, Tasumi, M, Tanokurab, M, Hiraoki, T, Yazawa, M & Tsutsumi, A 1994 Infrared studies of interaction between metal ions and Ca'-binding proteins. FEBS Letters 349 84–88 CrossRefGoogle Scholar
Nonaka, M, Li-Chan, E & Naki, S 1993 Raman spectroscopic study of thermally induced gelation of whey proteins. Journal of Agricultural and Food Chemical 41 1176–1181 CrossRefGoogle Scholar
Rajagapol, K & Schneider, JP 2004 Self-assembling peptides and proteins for nanotechnological applications. Current Opinion in Structural Biology 14 480–486 CrossRefGoogle Scholar
Rava, RP & Spiro, TG 1984 Ultraviolet resonance Raman spectra of insulin and α-lactalbumin with 218-and 200-nm laser excitation. Biochemistry 24 1861–1865 CrossRefGoogle Scholar
Spiro, TG & Gaber, BP 1977 Laser Raman scattering as a probe of protein structure. Annual Review of Biochemistry 46 553–572 CrossRefGoogle ScholarPubMed
Tu, AT 1986 Peptide backbone conformation and microenvironment of protein side chains in spectroscopy of biological systems In: Spectroscopy of Biological Systems, pp. 47–112 (Eds Clark, RJH & Haster, RE) New York: John Wiley & Sons Google Scholar
Wilson, G, Ford, SJ, Cooper, A, Hecht, L, Wen, ZQ & Barron, LD 1995 Vibrational Raman optical activity of alpha-lactalbumin: comparison with lysozyme, and evidence for native tertiary folds in molten globule states. Journal of Molecular Biology 254 747–760 CrossRefGoogle ScholarPubMed
Xie, M-X & Liu, Y 2003 Studies on amide III infrared bands for the secondary structure determination of proteins. Chemical Journal of Chinese Universities 24 226–231 Google Scholar
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