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Originally published In Press as doi:10.1074/mcp.M500085-MCP200 on April 25, 2005.
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Molecular & Cellular Proteomics 4:984-992, 2005.
© 2005 by The American Society for Biochemistry and Molecular Biology, Inc.


Research

Identification of Placental Transforming Growth Factor-ß and Bikunin Metabolites as Contaminants of Pharmaceutical Human Chorionic Gonadotrophin Preparations by Proteomic Techniques*

Sotiris Malatos{ddagger},§, Hendrik Neubert,||, Andrew T. Kicman,** and Ray K. Iles{ddagger},§,{ddagger}{ddagger}

From the {ddagger} Biomedical Sciences, Institute of Social and Health Research, Middlesex University, Enfield EN3 4SA, United Kingdom, § The Williamson Laboratory, St. Bartholomew’s and the Royal London School of Medicine and Dentistry, Queen Mary University of London, London EC1A 7BE, United Kingdom, and Drug Control Centre, King’s College London, London SE1 9NH, United Kingdom

A contaminant protein complex found in pharmaceutical urinary human chorionic gonadotrophin preparations is reported to have anti-human immunodeficiency virus-associated Kaposi’s sarcoma activity. The aim of this study was to isolate and characterize this protein complex by proteomic approaches. Size exclusion chromatography was used in the isolation of these human chorionic gonadotrophin-associated fragments. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the presence of a protein complex that dissociated into two protein bands under reducing conditions. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry of this complex showed three polypeptides at ~6.2, 11.4, and 15.8 kDa. Peptide mass mapping and N-terminal amino acid sequencing identified two polypeptides as metabolites of placental transforming growth factor-ß (11.4 kDa) and bikunin (15.8 kDa). Subsequent matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of the anti-human immunodeficiency virus-associated Kaposi’s sarcoma active preparations CG-10 (Sigma), Pregnyl (Organon), and Profasi (Serono) revealed the presence of metabolites of placental transforming growth factor-ß in all three; no other non-human chorionic gonadotrophin-related protein species were observed in these preparations. Our findings present evidence that urinary human chorionic gonadotrophin preparations are contaminated with metabolites of placental transforming growth factor-ß, which may have transforming growth factor-ß agonist actions, and metabolites of bikunin, which is a protease inhibitor. In combination these molecules may be responsible for the anti-human immunodeficiency virus-associated Kaposi’s sarcoma activity demonstrated for these urinary human chorionic gonadotrophin preparations.


** To whom correspondence may be addressed. E-mail: andrew.kicman{at}kcl.ac.uk.

{ddagger}{ddagger} To whom correspondence may be addressed: Biomedical Sciences, Inst. of Social and Health Research, Middlesex University, Enfield EN3 4SA, UK. E-mail: r.iles{at}mdx.ac.uk.


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eLetters:

Read all eLetters

Human chorionic gonadotropin and Kaposi’s sarcoma
Thierry Simonart
MCP Online, 12 Jun 2005 [Full text]
TGF{beta} receptor antagonism may explain urinary hCG effects on Kaposi's sarcoma growth
Ray Iles
MCP Online, 13 Jun 2005 [Full text]



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