Study on the secondary structure and hydration effect of human serum albumin under acidic pH and ethanol perturbation with IR/NIR spectroscopy
Abstract
Human serum albumin (HSA) is the most abundant protein in plasma and plays an essential physiological role in the human body. Ethanol precipitation is the most widely used way to obtain HSA, and pH and ethanol are crucial factors affecting the process. In this study, infrared (IR) spectroscopy and near-infrared (NIR) spectroscopy in combination with chemometrics were used to investigate the changes in the secondary structure and hydration of HSA at acidic pH (5.6–3.2) and isoelectric pH when ethanol concentration was varied from 0% to 40% as a perturbation. IR spectroscopy combined with the two-dimensional correlation spectroscopy (2DCOS) analysis for acid pH system proved that the secondary structure of HSA changed significantly when pH was around 4.5. What’s more, the IR spectroscopy and 2DCOS analysis showed different secondary structure forms under different ethanol concentrations at the isoelectric pH. For the hydration effect analysis, NIR spectroscopy combined with the McCabe–Fisher method and aquaphotomics showed that the free hydrogen-bonded water fluctuates dynamically, with ethanol at 0–20% enhancing the hydrogen-bonded water clusters, while weak hydrogen-bonded water clusters were formed when the ethanol concentration increased continuously from 20% to 30%. These measurements provide new insights into the structural changes and changes in the hydration behavior of HSA, revealing the dynamic process of protein purification, and providing a theoretical basis for the selection of HSA alcoholic precipitation process parameters, as well as for further studies of complex biological systems.
References
- 1. , Structural and Functional Modification of Human Serum Albumin by Lipid Peroxidation By-Products (Duquesne University ProQuest Dissertations Publishing, USA, 2005). Google Scholar
- 2. , “Structure of serum-albumin,” Adv. Protein Chem. 45, 153–203 (1994). Crossref, Web of Science, Google Scholar
- 3. , “All about albumin,” All About Albumin 319–413 (1995). Google Scholar
- 4. J. R. Brown, “Structure of serum-albumin,” Abs. Papers Am. Chem. Soc. 172(Sep3), 12 (1976). Google Scholar
- 5. , “Chemical, clinical, and immunological studies on the products of human plasma fractionation. I. The characterization of the protein fractions of human plasma,” J. Clin. Invest. 23(4), 417–432 (1944). Crossref, Google Scholar
- 6. , “Investigation of protective effect of ethanol on the natural structure of protein with infrared spectroscopy,” Spectrochim. Acta A-Mol. Biomol. Spectrosc. 271, 120935 (2022). Crossref, Web of Science, Google Scholar
- 7. , “Biomolecular and bioanalytical applications of infrared spectroscopy — A review,” Anal. Chim. Acta. 1133, 150–177 (2020). Crossref, Web of Science, Google Scholar
- 8. , “Near-infrared spectroscopy for in-line monitoring of protein unfolding and its interactions with lyoprotectants during freeze-drying,” Anal. Chem. 84(2), 947–955 (2012). Crossref, Web of Science, Google Scholar
- 9. , “Modeling temperature-dependent protein structural transitions by combined near-IR and mid-IR spectroscopies and multivariate curve resolution,” Anal. Chem. 75(20), 5592–5601 (2003). Crossref, Web of Science, Google Scholar
- 10. , “Investigation of water interaction with polymer matrices by near-infrared (NIR) spectroscopy,” Molecules 27(18), 5882 (2022). Crossref, Web of Science, Google Scholar
- 11. , “Determination of the immunoglobulin G precipitation end-point by an intelligent near-infrared spectroscopy system,” J. Innov. Opt. Health Sci. 14(3), 2150007 (2021). Link, Web of Science, Google Scholar
- 12. , “Investigating the structural change in protein aqueous solution using temperature-dependent near-infrared spectroscopy and continuous wavelet transform,” Appl. Spectrosc. 71(3), 472–479 (2017). Crossref, Web of Science, Google Scholar
- 13. , “Understanding the function of water during the gelation of globular proteins by temperature-dependent near infrared spectroscopy,” Phys. Chem. Chem. Phys. 20(30), 20132–20140 (2018). Crossref, Web of Science, Google Scholar
- 14. , Two-Dimensional Nuclear Magnetic Resonance in Liquids, Delft University Press, USA and Canada (1982). Google Scholar
- 15. , “Two-dimensional vibrational circular dichroism correlation spectroscopy: pH-induced spectral changes in L-alanine,” J. Mol. Struct. 799(1–3), 226–238 (2006). Crossref, Web of Science, Google Scholar
- 16. , Visible-near infrared perturbation spectroscopy: Water in action seen as a source of information, 12th Int. Conf. Near-Infrared Spectroscopy,
Auckland (2005), pp. 607–612. Google Scholar - 17. , “Application of near-infrared spectroscopy to agriculture and food analysis,” Guang Pu Xue Yu Guang Pu Fen Xi 24(4), 447–450 (2004). Google Scholar
- 18. , “Quantification of potassium concentration with Vis/SWNIR spectroscopy in fresh lettuce,” J. Innov. Opt. Health Sci. 13(6), 2050029 (2020). Link, Web of Science, Google Scholar
- 19. , “Near-infrared spectroscopy applications in pharmaceutical analysis,” Talanta 72(3), 865–883 (2007). Crossref, Web of Science, Google Scholar
- 20. , “Multivariety and multimanufacturer drug identification based on near-infrared spectroscopy and recurrent neural network,” J. Innov. Opt. Health Sci. 15(4), 2250022 (2022). Link, Google Scholar
- 21. , “Rapid and simultaneous determination of moisture and berberine content in Coptidis Rhizoma and Phellodendri Chinensis Cortex by near-infrared spectroscopy and chemometrics,” J. Innov. Opt. Health Sci. 13(2), 2050006 (2020). Link, Web of Science, Google Scholar
- 22. , “Assessment of cerebral oxygenation response to hemodialysis using near-infrared spectroscopy (NIRS): Challenges and solutions,” J. Innov. Opt. Health Sci. 14(6), 2150016 (2021). Link, Google Scholar
- 23. , “Near-infrared spectroscopy as a promising tool in stroke: Current applications and future perspectives,” J. Innov. Opt. Health Sci. 14(6), 2130006 (2021). Link, Google Scholar
- 24. , “Spectra selection methods: A novel optimization way for treating dynamic spectra and in-line near infrared modelling,” J. Innov. Opt. Health Sci. 13(4), 2050015 (2020). Link, Web of Science, Google Scholar
- 25. , “Multi-manufacturer drug identification based on near infrared spectroscopy and deep transfer learning,” J. Innov. Opt. Health Sci. 13(4), 2050016 (2020). Link, Web of Science, Google Scholar
- 26. , “Aquaphotomics approach for monitoring different steps of purification process in water treatment systems,” Talanta 206, 120253 (2020). Crossref, Web of Science, Google Scholar
- 27. , “Development of calibration models for rapid determination of moisture content in rubber sheets using portable near-infrared spectrometers,” J. Innov. Opt. Health Sci. 13(2), 2050009 (2020). Link, Web of Science, Google Scholar
- 28. , “Correlation between chain architecture and hydration water structure in polysaccharides,” Biomacromolecules 17(3), 1198–1204 (2016). Crossref, Web of Science, Google Scholar
- 29. , Near-Infrared Spectroscopy: Theory, Spectral Analysis, Instrumentation, and Applications, Springer, Singapore (2021). Crossref, Google Scholar
- 30. , “Two-dimensional infrared spectroscopy and principle component analysis studies of the secondary structure and kinetics of hydrogen-deuterium exchange of human serum albumin,” J. Phys. Chem. B 105(26), 6251–6259 (2001). Crossref, Web of Science, Google Scholar
- 31. , “Infrared spectroscopy of hydrogen-bonding interactions in neutral dimethylamine-methanol complexes,” J. Phys. Chem. A 123(46), 10109–10115 (2019). Crossref, Web of Science, Google Scholar
- 32. , “A new possibility of the generalized two-dimensional correlation spectroscopy. 1. Sample-sample correlation spectroscopy,” J. Phys. Chem. A 104(27), 6380–6387 (2000). Crossref, Web of Science, Google Scholar
- 33. , “Perturbation-correlation moving-window two-dimensional correlation spectroscopy,” Appl. Spectrosc. 60(4), 398–406 (2006). Crossref, Web of Science, Google Scholar
- 34. , “Two-dimensional/attenuated total reflection infrared correlation spectroscopy studies on secondary structural changes in human serum albumin in aqueous solutions: pH-dependent structural changes in the secondary structures and in the hydrogen bondings of side chains,” J. Phys. Chem. B 105(20), 4763–4769 (2001). Crossref, Web of Science, Google Scholar
- 35. ,
ScienceDirect , All About Albumin: Biochemistry, Genetics, and Medical Applications, Academic Press, San Diego (1996). Google Scholar - 36. , “Structure of serum albumin,” Adv. Protein Chem. 45, 153–203 (1994). Crossref, Web of Science, Google Scholar
- 37. , “Cd-resolved secondary structure of bovine plasma-albumin in acid-induced isomerization,” Int. J. Peptide Protein Res. 22(3), 333–340 (1983). Crossref, Google Scholar
- 38. , Two-Dimensional Correlation Spectroscopy: Applications in Vibrational and Optical Spectroscopy, Wiley, Hoboken (2005). Google Scholar
- 39. , “Concatenated two-dimensional correlation analysis: A new possibility for generalized two-dimensional correlation spectroscopy and its application to the examination of process reversibility,” Appl. Spectrosc. 64(3), 343–350 (2010). Crossref, Web of Science, Google Scholar
- 40.
L. Weyer , Practical Guide to Interpretive Near-Infrared Spectroscopy, CRC Press, Boca Raton (2008). Google Scholar - 41. , “Research on the structure of peanut allergen protein ara h1 based on aquaphotomics,” Front. Nutr. 8, 696355 (2021). Crossref, Web of Science, Google Scholar
- 42. , “Near-infrared spectroscopic method for investigating the hydration of a solute in aqueous solution,” J. Phys. Chem. 74(15), 2990–2998 (1970). Crossref, Web of Science, Google Scholar
- 43. , “Analysis of hydration water around human serum albumin using near-infrared spectroscopy,” Int. J. Biol. Macromol. 138, 927–932 (2019). Crossref, Web of Science, Google Scholar
- 44. , “Aquaphotomics: dynamic spectroscopy of aqueous and biological systems describes peculiarities of water,” J. Near Infrared Spectrosc. 17(6), 303–313 (2009). Crossref, Web of Science, Google Scholar
- 45. , “Water confined in the local field of ions,” Chem. Phys. Chem. 15(18), 4077–4086 (2014). Crossref, Google Scholar
- 46. , “Use of near infrared hyperspectral imaging to identify water matrix co-ordinates in mushrooms (Agaricus bisporus) subjected to mechanical vibration,” J. Near Infrared Spectrosc. 17(6), 363–371 (2009). Crossref, Web of Science, Google Scholar
- 47. , “Understanding hyaluronic acid induced variation of water structure by near-infrared spectroscopy,” Sci. Rep. 10(1), 1–8 (2020). Web of Science, Google Scholar
- 48. , “Water molecular system dynamics associated with amyloidogenic nucleation as revealed by real time near infrared spectroscopy and aquaphotomics,” PLoS One 9(7), e101997 (2014). Crossref, Web of Science, Google Scholar