Abstract
We have studied the effect of 2,2,2-trifluoroethanol (TFE), an α-helix inducer, versus methyl cyanide (MeCN), a β-sheet inducer, on acid-denatured human serum albumin (HSA) using far-UV circular dichroism, intrinsic fluorescence, 1-anilino-8-naphthalene sulfonate binding, and acrylamide quenching studies. Interestingly, at pH 2.0, where the recovery and resolution of the protein in reverse phase chromatography is high, its secondary structure remains unchanged even in the presence of very high concentration (76% v/v) of MeCN. Gain of 23 and 34% α-helicity was observed in the presence of 20 and 50% TFE, respectively. At pH 7.3, HSA aggregates in the presence of 40% MeCN, but it remains soluble up to 75% MeCN at pH 2.0. The results seem to be important for HSA isolation and purification.
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Abbreviations
- ANS:
-
1-anilino-8-naphthalene sulfonic acid
- GuHCl:
-
guanidine hydrochloride
- HSA:
-
human serum albumin
- MeCN:
-
methyl cyanide (or acetonitrile)
- MRE:
-
mean residue ellipticity
- RFI:
-
relative fluorescence intensity
- RPC:
-
reverse phase chromatography
- TFE:
-
2,2,2-trifluoroethanol
- UV-CD:
-
ultraviolet circular dichroism
References
Anfinsen, C. B. (1973) Science, 181, 223–230.
Haq, S. K., Rasheedi, S., Sharma, P., Ahmad, B., and Khan, R. H. (2005) Int. J. Biochem. Cell Biol., 37, 361–374.
Bohidar, H. B., and Mohanty, B. (2004) Phys. Rev. E Stat. Nonlin. Soft Matter Phys., 69, 021902.
Peters, T., Jr. (1996) in All About Albumin: Biochemistry, Genetics, and Medical Applications, Academic Press, N. Y., pp. 251–284.
Peters, T., Jr. (1996) in All About Albumin: Biochemistry, Genetics, and Medical Applications, Academic Press, N. Y., pp. 9–54.
Muzammil, S., Kumar, Y., and Tayyab, S. (1999) Eur. J. Biochem., 266, 26–32.
Khan, F., Khan, R. H., and Muzammil, S. (2000) Biochim. Biophys. Acta, 1481, 229–236.
Buck, M., Radford, S. E., and Dobson, C. M. (1993) Biochemistry, 32, 669–678.
Fan, P., Bracken, C., and Baum, J. (1993) Biochemistry, 32, 1573–1582.
Kandori, K., Uoya, Y., and Ishikawa, T. (2002) J. Colloid Interface Sci., 252, 269–275.
Gekko, K., Ohmae, E., Kameyama, K., and Takagi, T. (1998) Biochim. Biophys. Acta, 1387, 195–205.
Kumar, Y., Muzammil, S., and Tayyab, S. (2005) J. Biochem., 138, 335–341.
Andrade, M. A., Chacon, P., Merelo, J. J., and Moran, F. (1993) Protein Eng., 6, 383–390.
Eftink, M. R., and Ghiron, C. A. (1976) Biochemistry, 15, 672–680.
Naeem, A., Khan, K. A., and Khan, R. H. (2004) Arch. Biochem. Biophys., 432, 79–87.
Ahmad, B., Khan, M. K. A., Haq, S. K., and Khan, R. H. (2004) Biochem. Biophys. Res. Commun., 314, 166–173.
Rasheed, Z., Khan, M. W., and Ali, R. (2006) Autoimmunity, 39, 479–488.
Kundu, S., Sundd, M., and Jagannadham, M. V. (2002) J. Biochem. Mol. Biol., 35, 155–164.
Matulis, D., Baumann, C. G., Bloomfield, V. A., and Lovrien, R. E. (1999) Biopolymers, 49, 451–458.
Ahmad, B., Ankita, and Khan, R. H. (2005) Arch. Biochem. Biophys., 437, 159–167.
Bhakuni, V. (1998) Arch. Biochem. Biophys., 357, 274–284.
He, X. M., and Carter, D. C. (1992) Nature, 358, 209–215.
Manavalan, P., and Johnson, W. C. (1983) Nature, 305, 831–832.
Pittz, E. P., and Timasheff, S. N. (1978) Biochemistry, 17, 615–623.
Dobson, C. M. (2003) Nature, 426, 884–890.
Ohnishi, S., and Takano, K. (2004) Cell Mol. Life Sci., 61, 511–524.
Srisailam, S., Kumar, T. K., Srimathi, T., and Yu, C. (2002) J. Am. Chem. Soc., 124, 1884–1888.
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Published in Russian in Biokhimiya, 2010, Vol. 75, No. 3, pp. 447–456.
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Sen, P., Iqbal, M.A., Fatima, S. et al. Methyl cyanide induces α to β transition and aggregation at high concentrations in E-state of human serum albumin. Biochemistry Moscow 75, 367–374 (2010). https://doi.org/10.1134/S0006297910030132
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DOI: https://doi.org/10.1134/S0006297910030132