Development of methods for comparative proteomic uroanalysis in pregnant and nonpregnant women to examine the processes occurring during pregnancy


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Abstract

Objective. To obtain experimental data on urinary proteome in women with physiological pregnancy in order to enhance the diagnostic value of urinary markers for different diseases. Subjects and methods. Urine was sampled from 10 non-pregnant women aged 24 to 34 years to standardize a urine collection protocol. The investigators chose a second morning specimen as an object for urinalysis and 20 minutes as the recommended time for the ice storage of the object before pretreatment. For the standardization of the urine sampling protocol for pregnant women to make a further proteomic analysis, urine specimens were taken from 10 women aged 23 to 35 years at 15-17 weeks of normal pregnancy with the total urinary protein quantity of no more than 100 μg/ml (no proteinuria). The urinary specimens were analyzed using an Agilent 1100 nanoflow high-performance liquid chromatograph (nano-HPLC) (Agilent, USA) and a Finnigan LTQ FT mass-spectrometer (ThermoElectron, Bremen, Germany). Results. A protocol to isolate protein extracts from urine specimens, which involves a procedure for primary urine sampling from female patients and extraction of protein mixtures from the urine specimens, was developed and standardized. This investigation showed the high reproducibility and informative value of urinary proteome data in the pregnant women, which allows us to propose this protocol to determine pregnancy-associated urinary proteomic changes and characteristic qualitative ones in the urine protein profile during pregnancy and to investigate whether the found protein molecules might be used as biomarkers for the early diagnosis of different pregnancy-related diseases.

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About the authors

N. L Starodubtseva

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia; Institute for Energy Problems of Chemical Physics, Russian Academy of Sciences

Email: proteom.oparina4@gmail.com
PhD, Head of the Laboratory of Proteomics of Human Reproduction; scientist of Laboratory of Ion and Molecular Physics

O. V Vavina

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: o_vavina@oparina4.ru
obstetrician-gynecologist, maternity department

Z. S Khodzhaeva

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: z_khodzhaeva@oparina4.ru
MD, Professor, Principal Investigator, Department of Maternal-Fetal Medicine

A. S Kononikhin

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia; Institute for Energy Problems of Chemical Physics, Russian Academy of Sciences

Email: konoleha@yandex.ru
Cand., a researcher at the Laboratory of Proteomics of Human Reproduction; Senior Research Fellow Laboratory of ionic and molecular physics

I. A Popov

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia; N.M. Emanuel Institute of Biochemical Physics, Russian Academy of Sciences

Email: hexapole@gmail.com
Cand., Senior Scientist, Laboratory of mass spectrometry of biomacromolecules

A. E Bugrova

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia; N.M. Emanuel Institute of Biochemical Physics, Russian Academy of Sciences

Senior Scientist, Laboratory of Neurochemistry

Z. I Usmanova

Institute for Energy Problems of Chemical Physics, Russian Academy of Sciences; Moscow Institute of Physics and Technology

Email: ziliya.usmanova@gmail.com
Students, Laboratory of ionic and molecular physics

Yu. I Kostyukevich

Institute for Energy Problems of Chemical Physics, Russian Academy of Sciences; Moscow Institute of Physics and Technology

Email: yura542@gmail.com
Researcher, Laboratory of ionic and molecular physics

M. Yu Vysokikh

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: m_vysokikh@oparina4.ru
PhD, Head of mitochondrial medicine research group

N. E Kan

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: n_kan@oparina4.ru
MD, chief physician

E. N Nikolaev

Institute for Energy Problems of Chemical Physics, Russian Academy of Sciences; N.M. Emanuel Institute of Biochemical Physics, Russian Academy of Sciences

Email: ennikolaev@rambler.ru
prof., the head of Laboratory of Mass Spectrometry of Biomacromolecules

G. T Sukhikh

Academician V.I. Kulakov Research Center of Obstetrics, Gynecology and Perinatology, Ministry of Health of Russia

Email: g_sukhikh@oparina4.ru
MD, PhD, Professor, Director

References

  1. Schaub S., Wilkins J., Weiler T., Sangster K., Rush D., Nickerson P. Urine protein profiling with surface-enhanced laser-desorption/ionization time-of-flight mass spectrometry. Kidney Int. 2004; 65(1): 323-32.
  2. Theodorescu D., Wittke S., Ross M.M., Walden M., Conaway M., Just I. et al. Discovery and validation of new protein biomarkers for urothelial cancer: a prospective analysis. Lancet Oncol. 2006; 7(3): 230-40.
  3. Candiano G., Musante L., Bruschi M., Petretto A., Santucci L., Del Boccio P. et al. Repetitive fragmentation products of albumin and alphai-antitrypsin in glomerular diseases associated with nephrotic syndrome. J. Am. Soc. Nephrol. 2006; 17(11): 3139-48.
  4. Fliser D., Wittke S., Mischak H. Capillary electrophoresis coupled to mass spectrometry for clinical diagnostic purposes. Electrophoresis. 2005; 26: 2708-16.
  5. Mischak H., Apweiler R., Banks R.E., Conaway M., Coon J., Dominiczak A. et al. Clinical proteomics: A need to define the field and to begin to set adequate standards. Proteomics Clin. Appl. 2007; 1(2): 148-56.
  6. Pisitkun T., Shen R.F., Knepper M.A. Identification and proteomic profiling of exosomes in human urine. Proc. Natl. Acad. Sci. USA. 2004; 101: 13368-73.
  7. Saetun P., Semangoen T., Thongboonkerd V. Characterizations of urinary sediments precipitated after freezing and their effects on urinary pro tein and chemical analyses. Am. J. Physiol. Renal Physiol. 2009; 296: F1346-54.
  8. Агрон И.А., Автономов Д.М., КононихинА.С., Попов И.А., Мошковский С.А., Николаев Е.Н. База данных по точным массово-временным меткам для хромато-масс-спектрометрического анализа протеома мочи. Биохимия. 2010; 25(4): 598-605. [Agron I.A., Avtonomov D.M., Kononikhin A.S., Popov I.A., Moshkovsky S.A., Nikolaev E.N. Database of accurate mass-timestamp for gas chromatography-mass spectrometry analysis of urine proteome. Biochemistry. 2010; 25 (4): 598-605. In Russ.]
  9. Стародубцева Н.Л., Попов И.А., Николаев Е.Н., Иванец Т.Ю., Алексеева М.Л., Логинова Н.С., Николаева А.В., Вавина О.В., Сухих Г.Т. Поиск воспроизводимых биомаркеров для диагностики преэклам-псии. Акушерство и гинекология. 2013; 2: 10-7. [Starodubtseva N.L., Popov I.A., Nikolaev E.N., Ivanets T.Yu., Alekseeva M.L., Loginova N.S., Nikolaeva A.V., Vavina O.V., Sukhikh G.T. Search of reproducible biomarkers for the diagnosis of pre-eclampsia. Obstetrics and gynecology. 2013; 2: 10-7. In Russ.]
  10. Conrads T.P., Anderson G.A., Veenstra T.D., Pasa-Tolic L., Smith R.D. Utility of accurate mass tags for proteome-wide protein identification. Anal. Chem. 2000; 72(14): 3349-54.
  11. Belov M.E., Rakov V.S., Nikolaev E.N., Goshe M.B., Anderson G.A., Smith R.D. Initial implementation of external accumulation liquid chromatography/ electrospray ionization Fourier transform ion cyclotron resonance with automated gain control. Rapid Commun. Mass Spectrom. 2003; 17(7): 627-36.
  12. Shen Y., Tolić N., Masselon C., Pasa-Tolić L., Camp D.G. 2nd, Hixson K.K. et al. Ultrasensitive proteomics using high-efficiency on-line micro-SPE-nanoLC-nanoESI MS and MS/MS. Anal. Chem. 2004; 76(1): 144-54.
  13. Zubarev R.A., Hakansson P., Sundqvist B. Accuracy requirements for peptide characterization by monoisotopic molecular mass measurements. Anal. Chem. 1996; 68: 4060-3.
  14. Song Q., Chao J., Chao L. High level expression of human tissue kallikrein in the circulation induces hypotension in transgenic mice. Immunopharmacology. 1996; 32: 105-7.
  15. Carretero O.A. Vascular remodeling and the kallikrein-kinin system. J. Clin. Invest. 2005; 115: 588-91.
  16. Thebault S., Alexander R.T., Tiel Groenestege W.M., Hoenderop J.G., Bindels R.J. EGF increases TRPM6 activity and surface expression. J. Am. Soc. Nephrol. 2009; 20: 78-85.
  17. Christensen B., Petersen T.E., Sorensen E.S. Post-translational modification and proteolytic processing of urinary osteopontin. Biochem. J. 2008; 411: 53-61.
  18. Schmid M., Prajczer S., Gruber L.N., Bertocchi C., Gandini R., Pfaller W. et al. Uromodulin facilitates neutrophil migration across renal epithelial monolayers. Cell. Physiol. Biochem. 2010; 26(3): 311-8.
  19. Barratt J., Topham P. Urine proteomics: the present and future of measuring urinary protein components in disease. Can. Med. Assoc. J. 2007; 177: 361-8.

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