Export file:

Format

  • RIS(for EndNote,Reference Manager,ProCite)
  • BibTex
  • Text

Content

  • Citation Only
  • Citation and Abstract

Universal buffers for use in biochemistry and biophysical experiments

Department of Chemistry and Biochemistry, Montana State University, Bozeman MT 59717, USA

The use of buffers that mimic biological solutions is a foundation of biochemical and biophysical studies. However, buffering agents have both specific and nonspecific interactions with proteins. Buffer molecules can induce changes in conformational equilibria, dynamic behavior, and catalytic properties merely by their presence in solution. This effect is of concern because many of the standard experiments used to investigate protein structure and function involve changing solution conditions such as pH and/or temperature. In experiments in which pH is varied, it is common practice to switch buffering agents so that the pH is within the working range of the weak acid and conjugate base. If multiple buffers are used, it is not always possible to decouple buffer induced change from pH or temperature induced change. We have developed a series of mixed biological buffers for protein analysis that can be used across a broad pH range, are compatible with biologically relevant metal ions, and avoid complications that may arise from changing the small molecule composition of buffers when pH is used as an experimental variable.
  Figure/Table
  Supplementary
  Article Metrics

References

1. Pascal SM, Yamazaki T, Singer AU, et al. (1995) Structural and dynamic characterization of the phosphotyrosine binding region of a Src homology 2 domain--phosphopeptide complex by NMR relaxation, proton exchange, and chemical shift approaches. Biochemistry 34:11353-11362.    

2. Zhang M, Zhou M, van Etten RL, et al. (1997) Crystal structure of bovine low molecular weight phosphotyrosyl phosphatase complexed with the transition state analog vanadate. Biochemistry 36:15-23.    

3. Katayama DS, Nayar R, Chou DK, et al. (2006) Effect of buffer species on the thermally induced aggregation of interferon-tau. J Pharm Sci 95:1212-1226.    

4. Hausamen TU, Helger R, Rick W, et al. (1967) Optimal conditions for determination of serum alkaline phosphatase by a new kinetic method. Clin Chim Acta 15: 241-245.    

5. McComb RB, Bowers GN (1972) Study of optimum buffer conditions for measuring alkaline-phosphatase activity in human serum. Clin Chem 18: 97-103.

6. Ganichkin OM, Xu XM, Carlson BA, et al. (2008) Structure and catalytic mechanism of eukaryotic selenocysteine synthase. J Biol Chem 283: 5849-5865.

7. Long D, Yang D (2009) Buffer Interference with Protein Dynamics: A Case Study on Human Liver Fatty Acid Binding Protein. Biophys J 96 1482-1488.

8. Sopkova J, Renouard M, Lewit-Bentley A (1993) The crystal structure of a new high-calcium form of annexin V. J Mol Biol 234: 816-825.    

9. Kumar S, Sharma P, Arora K, et al. (2014) Calcium binding to beta-2-microglobulin at physiological pH drives the occurrence of conformational changes which cause the protein to precipitate into amorphous forms that subsequently transform into amyloid aggregates. PLoS One 9: e95725.    

10. Schmitt FJ, Thaa B, Junghans C, et al. (2014) eGFP-pHsens as a highly sensitive fluorophore for cellular pH determination by fluorescence lifetime imaging microscopy (FLIM). Biochim Biophys Acta 1837: 1581-1593.    

11. Bothner B, Schneemann A, Marshall D, et al. (1999) Crystallographically identical virus capsids display different properties in solution. Nat Struct Bio 6: 114-116.    

12. Bothner B, Taylor D, Jun B, et al. (2005) Maturation of a tetravirus capsid alters the dynamic properties and creates a metastable complex. Virology 339: 145-145.    

13. Nam HJ, Gurda BL, McKenna R, et al. (2011) Structural Studies of Adeno-Associated Virus Serotype 8 Capsid Transitions Associated with Endosomal Trafficking. J Virol 85: 11791-11799.    

14. Bartlett JS, Wilcher R, Samulski RJ (2000) Infectious entry pathway of adeno-associated virus and adeno-associated virus vectors. J Virol 74: 2777-2785.    

15. Roos WH, Bruinsma R, Wuite GJL (2010) Physical virology. Nat Phys 6: 733-743.    

16. Ellis DA (1961) New universal buffer system. Nature 191: 1099-1100.    

17. Henderson Y (1908) Acapnia and Shock: Carbon-dioxid as a factor in the regulation of the heart-rate. Am J Physiol 21: 126-156.

18. Good NE, Winget GD, Winter W, et al. (1966) Hydrogen ion buffers for biological research. Biochemistry 5: 467-472.    

19. Garrett RH, Charles M, Grisham CM (2012). Biochemistry 5th ed. Cengage Learning.

20. Sokolowska M, Bal J (2005) Cu(II) complexation by “non-coordinating” N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES buffer) J Inorg Biochem 99: 1653-1660.

21. Taha M (2011) Complex Equilibria in Aqueous Solutions of Chromium(III) with Some Biological pH Buffers. J Chem Eng Data 56: 3541-3551.    

22. Shnyrov VL, SanchezRuiz JM, Boiko BN, et al. (1997) Applications of scanning microcalorimetry in biophysics and biochemistry. Thermochim Acta 302: 165-180.    

23. Lakowicz J (2006) Principles of Fluorescence Spectroscopy. New York: Springer.

Copyright Info: © 2015, Brian Bothner, et al., licensee AIMS Press. This is an open access article distributed under the terms of the Creative Commons Attribution Licese (http://creativecommons.org/licenses/by/4.0)

Download full text in PDF

Export Citation

Article outline

Show full outline
Copyright © AIMS Press All Rights Reserved