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http://dx.doi.org/10.14348/molcells.2014.0214

Structural Identification of Modified Amino Acids on the Interface between EPO and Its Receptor from EPO BRP, Human Recombinant Erythropoietin by LC/MS Analysis  

Song, Kwang-Eun (Department of Stereoscopic Media, Korean German Institute of Technology)
Byeon, Jaehee (Department of Stereoscopic Media, Korean German Institute of Technology)
Moon, Dae-Bong (BIOnSYSTEMS, Inc., R&D Center)
Kim, Hyong-Ha (Center for Bioanalysis, Korea Research Institute of Standards and Science)
Choi, Yoo-Joo (Department of Newmedia, Korean German Institute of Technology)
Suh, Jung-Keun (Department of Newmedia, Korean German Institute of Technology)
Publication Information
Molecules and Cells / v.37, no.11, 2014 , pp. 819-826 More about this Journal
Abstract
Protein modifications of recombinant pharmaceuticals have been observed both in vitro and in vivo. These modifications may result in lower efficacy, as well as bioavailability changes and antigenicity among the protein pharmaceuticals. Therefore, the contents of modification should be monitored for the quality and efficacy of protein pharmaceuticals. The interface of EPO and its receptor was visualized, and potential amino acids interacting on the interface were also listed. Two different types of modifications on the interface were identified in the preparation of rHu-EPO BRP. A UPLC/Q-TOF MS method was used to evaluate the modification at those variants. The modification of the oxidized variant was localized on the Met54 and the deamidated variants were localized on the Asn47 and Asn147. The extent of oxidation at Met54 was 3.0% and those of deamidation at Asn47 and Asn147 were 2.9% and 4.8%, respectively.
Keywords
deamidation; erythropoietin; interface; modification; oxidation;
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1 Brines, M., Patel, N.S., Villa, P., Brines, C., Mennini, T., De Paola, M., Erbayraktar, Z., Erbayraktar, S., Sepodes, B., Thiemermann, C., et al. (2008). Nonerythropoietic, tissue-protective peptides derived from the tertiary structure of erythropoietin. Proc. Natl. Acad. Sci. USA 105, 10925-10930.   DOI   ScienceOn
2 Davis, J.M., Arakawa, T., Strickland, T.W., and Yphanitis, D.A. (1987). Characterization of recombinant human erythropoietin produced in Chinese hamster ovary cells. Biochemistry 26, 2633-2638.   DOI   ScienceOn
3 Chang, S.H., Kim, H.J., and Kim, C.W. (2013). Analysis of the structure and stability of erythropoietin by pH and temperature changes using various LC/MS. Bull. Korean Chem. Soc. 34, 2663-2670.   과학기술학회마을   DOI   ScienceOn
4 Cheetham, J.C., Smith, D.M., Aoki, K.H., Stevenson, J.L., Hoeffel, T.J., Syed, R.S., Egrie, J., and Harvey, T.S. (1998). NMR structure of human erythropoietin and a comparison with its receptor bound conformation. Nat. Struct. Biol. 5, 861-866.   DOI   ScienceOn
5 Delorme, E., Lorenzini, T., Giffin, J., Martin, F., Jacobsen, F., Boone, T., and Elliott, S. (1992). Role of glycosylation on the secretion and biological activity of erythropoietin. Biochemistry 31, 9871-9876.   DOI   ScienceOn
6 Elliott, S., Lorenzini, T., Chang, D., Barzilay, J., and Delorme, E. (1997). Mapping of the active site of recombinant human erythropoietin. Blood 89, 493-502.
7 Eschbach, J.W., Abdulhadi, M.H., Browne, J.K., Delano, B.G., Downing, M.R., Egrie, J.C., Evans, R.W., Friedman, E.A., Graber, S.E., Haley, N.R., et al. (1989). Recombinant human erythropoietin in anemic patients with end-stage renal disease. Results of a phase III multicenter clinical trial. Ann. Intern. Med. 111, 992-1000.   DOI   ScienceOn
8 Gan, Y., Xing, J., Jing, Z., Stetler, R.A., Zhang, F., Luo, Y., Ji, X., Gao, Y., and Cao, G. (2012). Mutant erythropoietin without erythropoietic activity is neuroprotective against ischemic brain injury. Stroke 43, 3071-3077.   DOI
9 Greer, F., Reason, A., and Rogers, M. (2002). Post-translational modifications of biopharmaceuticals-a challenge for analytical characterisation. Eur. Biopharm. Rev. 106-111.
10 Lodish, H.F., Hilton, D.J., Klingmuller, U., Watowich, S.S., and Wu, H. (1995). The erythropoietin receptor: biogenesis, dimerization, and intracellular signal transduction. Cold Spring Harb. Symp. Quant. Biol. 60, 93-104.
11 Syed, R.S., Reid, S.W., Li, C., Cheetham, J.C., Aoki, K.H., Liu, B., Zhan, H., Osslund, T.D., Chirino, A.J., Zhang, J., et al. (1998). Efficiency of signaling through cytokine receptors depends critically on receptor orientation. Nature 395, 511-516.   DOI   ScienceOn
12 Toyoda, T., Itai, T., Arakawa, T., Aoki, K.H., and Yamaguchi, H. (2000). Stabilization of human recombinant erythropoietin through interactions with the highly branched N-glycans. J. Biochem. 128, 731-737.   DOI   ScienceOn
13 Walsh, G. (2010). Post-translational modifications of protein biopharmaceuticals. Drug Discov. Today 15, 773-780.   DOI   ScienceOn
14 Winearls, C.G., Oliver, D.O., Pippard, M.J., Reid, C., Downing, M.R., and Cotes, P.M. (1986). Effect of human etythropoietin derived from recombinant DNA on the anaemia of patients maintained by chronic haemodialysis. Lancet 328, 1175-1178.   DOI   ScienceOn
15 Brinks, V., Hawe, A., Basmeleh, A.H., Joachin-Rodriguez, L., Haseberg, R., Somsen, G.W., Jiskoot, W., and Schellekens, H. (2011). Quality of original and biosimilar epoetin products. Pharm. Res. 28, 386-393.   DOI