Paper Titles in Periodical
International Letters of Chemistry, Physics and Astronomy
ILCPA Volume 38
Subscribe

Subscribe to our Newsletter and get informed about new publication regulary and special discounts for subscribers!

ILCPA > ILCPA Volume 38 > Mass Spectrometry Based Analysis of Erythrocyte...
Mass Spectrometry Based Analysis of Erythrocyte Membrane Associated Proteins in Chronic Myeloid Leukemia Patients in Sri Lanka
< Back to Volume

Mass Spectrometry Based Analysis of Erythrocyte Membrane Associated Proteins in Chronic Myeloid Leukemia Patients in Sri Lanka

Full Text PDF

Abstract:

The research reported in this paper was conducted to analyze erythrocyte membrane associated proteins (ERMBPs) of some of chronic myeloid leukemia (CML) patients and selected individuals of Sri Lanka employing one dimensional sodium dodecyl sulphate poly acrylamide gel electrophoresis (1D-SDS-PAGE) combined with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI–TOF-MS). Erythrocyte membranes from blood were isolated by osmotic lysis, centrifugation and washings. ERMBPs were separated on 1D-SDS-PAGE, visualized by silver staining and the separated protein bands dissected out from the gel and were subjected to digestion by proteolytic enzyme, trypsin, and the resulting peptide mixture was analyzed by MALDI-TOF-MS. Resulting experimental peptide mass values were analyzed by peptide mass fingerprint (PMF) technique. From this analysis 10 ERMBPs including α and β spectrin, ankyrin, band 3, band 4.1, band 4.2, band 7, dematin, actin, 55 KDa erythrocyte membrane protein were identified accurately with their primary structure information. The study was able to provide some evidence for Cathepsin associated cleavage of Band 3 anion transport protein in CML patients reported previously. In addition we were able to detect changes in gel bands between healthy controls and CML patients around the area of 20 kDa in the 1 D-SDS-PAGE. It was identified as nuclear protein Dbf 4 related factor 1 isoform 2. Although erythrocytes are devoid of nuclei, such unexpected nuclear proteins have been identified in previous research. We were successful in identifying several human ERMBPs with available resources. As the identified proteins were known to be related to pathology of some of the hematological diseases, this methodology could be extended to detect the protein changes in erythrocyte membrane protein associated diseases. Therefore, this initial research would at some point lead to discovery of biomarkers to these hematological diseases in Sri Lanka.

Info:

Periodical:
International Letters of Chemistry, Physics and Astronomy (Volume 38)
Pages:
74-86
DOI:
https://doi.org/10.18052/www.scipress.com/ILCPA.38.74
Citation:
D.U. Kottahachchi et al., "Mass Spectrometry Based Analysis of Erythrocyte Membrane Associated Proteins in Chronic Myeloid Leukemia Patients in Sri Lanka", International Letters of Chemistry, Physics and Astronomy, Vol. 38, pp. 74-86, 2014
Online since:
September 2014
Authors:
D.U. Kottahachchi, T.R. Ariyaratne, G.A.U. Jayasekera
Keywords:
1-D-SDS-PAGE, Erythrocyte Membrane Associated Proteins, Hematological Diseases, MALDI-TOF-MS, PMF
Export:
RIS, BibTeX, Text
Distribution:
Open Access

Creative Commons License
This work is licensed under a
Creative Commons Attribution 4.0 International License

References:

[1] M. Almén, K. J. Nordström, R. Fredriksson, H. B. Schiöth, BMC Biology 7 (2009) 50.

[2] M. E. Reid, N. Mohandas, Seminars in Hematology 41 (2004) 93-117.

[3] A. V. Hoffbrand, J. E. Pettit, P.A.H. Moss, Essential Haematology (Fourth Edition) (2004) Wiley-blackwell.

[4] E. M. Pasini, H.U. Lutz, M. Mann, A. W. Thomas Journal of proteomics 73 (2010b) 421-435.

[5] M. Kundu, J. Basu, M. Fujimagari, P. Williamson, R. A. Chlegel, P. Chakrabarti, BiochimBiophys Acta 1096(3) (1991) 205-8.

[6] J. Basu, M. Kundu, M. M. Rakshit, P. Chakrabarti, BiochimBiophysActa (1988) 121-126.

[7] R. Govekar, P. Kawle, R. Thomas, S. Advani, P. V. Sheena, S. Zingde, Anemia (2012) 1-7.

[8] H. Lahm, H. Langen, Electrophoresis 21 (2000) 2105-2114.

[9] O. Vorm, P. Roepstorff, M. Mann, Anal. Chem. 66 (1994) 3281-3287.

[10] S. J. Cottrell, Journal of Proteomics 74 (2011) 1842-1851.

[11] E. M. Pasini, M. Kirkegaard, P. Mortensen, H. U. Lutz, A. W. Thomas, M. Mann, Blood 108 (2006) 791-801.

[12] T.Y. Low, T.K. Seow, C.M. Chung, Proteomics 2 (2002) 1229-1239.

[13] S. Shattil, B. Furie, H. Cohen, L. Silberstein, Haematology: Basic Principals and Practice. Edinburgh, Churchill Livingstone, (1995) 672.

[14] M. M. B. Kay, S. R. Goodman, K. Sorensen, Proceedings of the National Academy of Sciences of the United States of America 80(6) (1983) 1631-1635.

[15] A. Santos-Silva, E. M. B. Castro, N. A. Teixeira, F. C. Guerra, A. Quintanilha, Clinica Chimica Acta 275(2) (1998) 185-196.

[16] E. M. Pasini, H. U. Lutz, M. Mann, A. W. Thomas, Journal of proteomics 73 (2010a) 403-420.

[17] F. Roux-Dalvai, A. Gonzalez de Peredo, C. Simo, L. Guerrier, D. Bouyssie, A. Zanella, A. Citterio, O. Burlet-Schiltz, E. Boschetti, P. G. Righetti, B. Monsarrat, Molecular & Cellular Proteomics 7(11) (2008) 2254-2269.

DOI: https://doi.org/10.1074/mcp.m800037-mcp200

[18] N. Yoshizawa-Sugata, A. Ishii, C. Taniyama, E. Matsui, K. Arai, H. Masai, The Journal of Biological Chemistry 280(13) (2005) 13062-13070.

DOI: https://doi.org/10.1074/jbc.m411653200

[19] A. Montagnoli, R. Bosotti, F. Villa, M. Rialland, D. Brotherton, C. Merurio, J. Berthelsen, Santocanale, C. The EMBO Journal 21(12) (2002) 3171-3181.

[20] E. M. Pasini, M. Kirkegaard, D. Salerno, P. Mortensen, M. Mann, A. W. Thomas, Molecular & Cellular Proteomics 7 (2008) 1317-1330. ( Received 12 August 2014; accepted 20 August 2014 ).

Show More Hide
Cited By:
This article has no citations.