JBPC  Vol.4 No.1 , February 2013
Protein and lipid characterization of wheat roots plasma membrane damaged by Fe and H2O2 using ATR-FTIR method
ABSTRACT

In plant cells the plasma membrane is a highly elaborated structure that functions as the point of exchange with adjoining cells, cell walls and the external environment. In this study, we investigated the structure and function characteristic of wheat root plasma membrane (PM) as affected by H2O2 and Fe by using fluorescence spectroscopic and attenuated total reflectance infrared (ATR-IR) techniques. The results showed that these oxidant damaged induced an obviously reduced membrane fluidity were observed in the roots PM treated with the 200 μM H2O2, FeSO4, and FeCl3. Computer-aided software analyses of the FTIR spectrum indicated that the content of the α-helices decreased and β-sheet increased in the secondary structures of proteins after exposure to the oxidants of 200 μM H2O2, FeSO4, and FeCl3. The number of P=O and C=C bonds area declined rapidly in the lipids of the membrane under the oxidants stress. These structural alterations might explain the reason of the roots PM instability under most of the abiotic stress.


Cite this paper
Zhao, X. , Yang, X. , Shi, Y. , Chen, G. and Li, X. (2013) Protein and lipid characterization of wheat roots plasma membrane damaged by Fe and H2O2 using ATR-FTIR method. Journal of Biophysical Chemistry, 4, 28-35. doi: 10.4236/jbpc.2013.41004.
References
[1]   Mittler, R. (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Science, 7, 5-10.

[2]   Davies, K.A., Sharon, W.L. and Pacifici, R.E. (1987) Protein damage and degradation by oxygen radicals. I. General aspects. Journal Biology Chemistry, 262, 9914-9920.

[3]   Stadtman, E.R. (1986) Oxidation of proteins by mixed-function oxidation systems: Implication in protein turnover, ageing and neutrophil function. Trends Biochemistry Science, 11, 11-12. doi:10.1016/0968-0004(86)90221-5

[4]   Wolff, S.P., Garner, A. and Dean, R.T. (1986) Free radicals, lipids and protein degradation. Trends Biochemistry Science, 11, 27-31. doi:10.1016/0968-0004(86)90228-8

[5]   Raven, J.A. (1990) Predictions of Mn and Fe use efficiencies of plant growth with different energy, carbon and nitrogen sources. New Phytologist, 109, 279-287. doi:10.1111/j.1469-8137.1988.tb04196.x

[6]   Ponnamperuma, F.N., Bradfield, J.F. and Peech, M. (1955) Physiological disease of rice attributable to iron toxicity. Nature, 175, 275. doi:10.1038/175265a0

[7]   Kneen, B.E., LaRue, T.A., Welch, R.M. and Weeden, N.F. (1990) Pleitropic effects of brz: A mutation in Pisum sativum (L.) cv “Sparke” conditioning decreased nodulation and increased ion uptake and leaf necrosis. Plant Physiology, 93, 717-723. doi:10.1104/pp.93.2.717

[8]   Briat, J.F., Forbis-Loisy, I., Grignon, N., Lobraux, S., Pascal, N., Savino, G., et al., (1995) Cellular and molecular aspects of iron metabolism in plants. Biology Cell, 84, 69-81. doi:10.1016/0248-4900(96)81320-7

[9]   Sills, R.H., Moore, D.J. and Mendelsohn, R. (1994) Erythrocyte peroxidation: Quantitation by fourier transform infrared spectroscopy. Analytical Biochemistry, 218, 118-123. doi:10.1006/abio.1994.1149

[10]   Yang, Y.L., Zhang, F., He, W.L., Wang, X.M. and Zhang L.X. (2003) Iron-mediated inhibition of H+-ATPase in plasma membrane vesicles isolated from wheat roots. Cellular and Molecular Life Sciences, 60, 1249-1257.

[11]   Goormaghtigh, E., Raussens, V. and Ruysschaert, J.M. (1999) Attenuated total re£ection infrared spectroscopy of proteins and lipids in biological membranes. Biochimica ET Biophysica Acta, 1422, 105-185. doi:10.1016/S0304-4157(99)00004-0

[12]   Phelan, A.M. and Lange, D.G. (1991) Ischemia/reperfusion-induced changes in membrane fluidity characteristics of brain capillary endothelial cells and its prevention by liposomal-incorporated superoxide dismutase. Biochemistry Biophysical Acta, 1067, 97. doi:10.1016/0005-2736(91)90030-C

[13]   Timasheff, S.N., Susi, H. and Stevens, L. (1967) Infrared spectra and protein conformations in aqueous solutions II. Survey of globular proteins. The Journal of Biological Chemistry, 242, 5467-5473.

[14]   Kasamo, K. and Sakibara, Y. (1995) The plasma membrane reconstitution into liposomes and its regulation by phospholipids. Plant Science, 111, 117-131. doi:10.1016/0168-9452(95)04224-I

[15]   Widell, S., Lundborg, T. and Christer, L. (1982) Plasma membranes from oats prepared by partition in an aqueous polymer two-phase system. Plant Physiology, 70, 1429-1435. doi:10.1104/pp.70.5.1429

[16]   Bradford, M. (1976) A Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. doi:10.1016/0003-2697(76)90527-3

[17]   Zhang, W.H., Chen, Q. and Liu, Y.L. (2002) Relationship between H+-ATPase activity and fluidity of tonoplast in barley roots under NaCl stress. Acta Botanical Sinica, 44, 292-296.

[18]   Witold, K., Surewicz, J., Mantsch, H. and Chapman, D. (1993) Determination of protein secondary structure by fourier transform infrared spectroscopy: A critical assessment? Biochemistry, 32, 19.

[19]   Zhao, X., Shi, Y., Chen, L., Sheng, F. and Zhou, H. (2011) Secondary structure changes and thermal stability of plasma membrane proteins of wheat roots in heat stress. American Journal of Plant Sciences, 2, 816-822. doi:10.4236/ajps.2011.26096

[20]   Byler, D.M. and Susi, H. (1986) Examination of the secondary structure of proteins by deconvolved FTIR spectra. Biopolymers, 25, 469-487. doi:10.1002/bip.360250307

[21]   Qiu, Q.S. and Su, X.F. (1998) The influence of extra cellular side Ca2+ on the activity of the plasma membrane H+-ATPase from wheat roots. Australian Journal Plant Physiology, 25, 923-928. doi:10.1071/PP98036

[22]   Qiu, Q.S. (1999) Influence of Osmotic stress on the lipid physical states of plasma membrane from wheat roots. Acta Botanical Sinica, 41, 161-165.

 
 
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