Renalase is a recently discovered secretory protein, which is suggested to play a role (which still remains elusive) in regulation of blood pressure. Earlier it was purified from urine of healthy volunteers by means of ammonium sulfate fractionation and subsequent affinity chromatography (Xu et al. (2005) J. Clin. Invest., 115, 1275). The resultant purified preparation of renalase contained 2 proteins with molecular masses of 35 and 67-75 kDa. The authors believed that the latter represents a dimerization (aggregation) product of the 35 kDa protein. In this study we have detected relanase in urinary samples of 2 of 6 volunteers only after immunoaffinity enrichment of urinary samples subjected to ammonium sulfate precipitation. Electrophoresis of the purified preparation also demonstrated the presence of 2 proteins with molecular masses of 35 and 66 kDa, respectively. Mass spectrometry analysis of these proteins identified 35 and 66 kDa proteins as renalase and serum albumin, respectively. Thus, our results do not support suggestion on formation of renalase dimers and they indicate that urinary renalase excretion significantly varies in humans.
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Keywords: renalase, urine, fractionation, immunoaffinity enrichment, mass spectrometry analysis
Citation:
Fedchenko V.I., Buneeva O.A., Kopylov A.T., Kaloshin A.A., Axenova L.N., Zgoda V.G., Medvedev A.E. (2012) Mass spectrometry detection of monomeric renalase in human urine. Biomeditsinskaya Khimiya, 58(5), 599-607.
Fedchenko V.I. et al. Mass spectrometry detection of monomeric renalase in human urine // Biomeditsinskaya Khimiya. - 2012. - V. 58. -N 5. - P. 599-607.
Fedchenko V.I. et al., "Mass spectrometry detection of monomeric renalase in human urine." Biomeditsinskaya Khimiya 58.5 (2012): 599-607.
Fedchenko, V. I., Buneeva, O. A., Kopylov, A. T., Kaloshin, A. A., Axenova, L. N., Zgoda, V. G., Medvedev, A. E. (2012). Mass spectrometry detection of monomeric renalase in human urine. Biomeditsinskaya Khimiya, 58(5), 599-607.
Xu J., Li G., Wang P., Velazquez H., Yao X., Li Y., Wu Y., Peixoto A., Crowley S., Desir G.V. (2005) J. Clin. Invest., 115, 1275-1280. CrossRef Scholar google search
Li G., Xu J., Wang P., Velazquez H., Li Y., Wu Y., Desir G.V. (2008) Circulation, 117, 1277-1282. CrossRef Scholar google search
Pandini V., Ciriello F., Tedeschi G., Rossoni G., Zanetti G., Aliverti A. (2010) Protein Expr. Purif., 72, 244-253. CrossRef Scholar google search
Milani M., Ciriello F., Baroni S., Pandini V., Canevari G., Bolognesi M., Aliverti A. (2011) J. Mol. Biol., 411, 463-473. CrossRef Scholar google search
Ghosh S.S., Krieg R.J., Sica D.A, Wang R., Fakhry I., Gehr T. (2009) Pediatr. Nephrol., 24, 367-377. CrossRef Scholar google search
Zhao Q., Fan Z., He J., Chen S., Li H., Zhang P., Wang L., Hu D., Huang J., Qiang B., Gu D. (2007) J. Mol. Med., 85, 877-885. CrossRef Scholar google search
Adachi J., Kumar Ch., Zhang Y., Olsen J.V., Mann M. (2006) Genome Biol., 9, R80. (http://genomebiology.com/2006/7/9/R80). Scholar google search
Hansen P.J. http://www.protocol-online.org/prot/Molecular_Biology/Protein/ Immunoprecipitation_IP_/index.html . Scholar google search
Doonan S.T. (ed.) (1996) In: Bulk Purification by Fractional Precipitation. New Jersey: Methods in Molecular Biology; pp. 135-150. Scholar google search
Mahn A., Ismail M. (2011) J. Chromatogr. B Analyt. Technol. Biomed. Life Sci., 879, 3645-3648. CrossRef Scholar google search
Soll D. (1995) tRNA: Structure, Biosynthesis and Function, ASM Press, New York. Scholar google search
Ward W.W., Swiatek G. (2011) Prot. Purif., Curr. Anal. Chem., 879, 3645-3648. Scholar google search
de Roos B., Duthie S.J., Polley A.C., Mulholland F., Bouwman F.G., Heim C., Rucklidge G.J., Johnson I.T., Mariman E.C., Daniel H., Elliott R.M. (2008) J. Proteome Res., 7, 2280-2290. CrossRef Scholar google search
