A possible cause of heterogeneity of mammalian apurinic/apyrimidinic endonuclease |
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| Institution: | 1. Division of Gastroenterology and Hepatology, Alameda Health System-Highland Hospital, Highland Care Pavilion, 5th floor, 1411 East 31st Street, Oakland, CA 94602, USA;2. Division of Gastroenterology and Hepatology, Stanford University School of Medicine, 750 Welch Road, Suite# 210, Palo Alto, CA 94304, USA;3. Liver Transplant Program, Stanford University Medical Center, 750 Welch Road, Suite# 210, Palo Alto, CA 94304, USA;4. Hepatitis B Foundation, 3805 Old Easton Road, Doylestown, PA 18902, USA;1. Barcelona Clinic Liver Cancer (BCLC) Group: Radiology Department. Hospital Clínic, University of Barcelona. CIBER ehd. Spain;2. Barcelona Clinic Liver Cancer (BCLC) Group: Liver Unit. Hospital Clínic, University of Barcelona. CIBER ehd. Spain;1. Barcelona Institute for Global Health (ISGlobal), Hospital Clínic, University of Barcelona, Spain;2. Department of Gastroenterology and Hepatology. Department of Medical and Health Sciences, Linköping University, Linköping, Sweden;3. Department of Medical & Surgical Sciences, “Alma Mater” University, Bologna, Italy;4. EASL International Liver Foundation, Geneva, Switzerland;5. University Medical Center Ljubljana, Dept. of Gastroenterology, Slovenia;6. Translational Research Group on Infectious Diseases of Lleida (TRIDLE), Infectious Diseases Clinical Direction, Biomedical Research Institute Dr Pifarré, University of Lleida, Lleida, Spain;7. Preventive Medicine and Epidemiology, Hospital Clínic, Barcelona, Spain;8. UCM Digestive Diseases, ciberehd and IBIS, Virgen del Rocío University Hospital, University of Seville, Seville, Spain;9. Pitie-Salpetriere Hospital, Department of Hepatology University Paris 6, France;10. Department of Hepatology/Gastroenterology, Charité University Medical Center, Berlin, Germany;11. Departamento de Gastrenterologia, CHLN, Clínica Universitária de Gastrenterologia, Laboratório de Nutrição, Faculdade de Medicina, Universidade de Lisboa, Portugal;12. Institute of Cellular Medicine, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, United Kingdom;13. The Liver Unit & NIHR Biomedical Research Centre, The Newcastle upon Tyne Hospitals NHS Foundation Trust, Newcastle upon Tyne, UK;1. UK Health Forum, London, United Kingdom;2. Departamento de Gastrenterologia, CHLN, Laboratório de Nutrição, Faculdade de Medicina, Universidade de Lisboa, Portugal;3. Divisions of Gastroenterology and Hepatology and Clinical Pathology, University Hospitals of Geneva, Geneva, Switzerland;4. Barcelona Institute for Global Health (ISGlobal), Hospital Clinic, University of Barcelona, Barcelona, Spain;5. CHIP, Rigshospitalet, University of Copenhagen, Øster Alle 56, 5. sal, DK-2100 Copenhagen, Denmark;6. University of Southampton, Southampton SO17 1BJ, United Kingdom;1. Université Paris-Saclay, Gustave Roussy, Villejuif, France;2. Memorial Sloan Kettering Cancer Center, New York;3. Weill Cornell College of Medicine, New York, USA;4. Trinity College Dublin, Dublin, Ireland;5. Mayo Clinic Cancer Center, Phoenix;6. Vanderbilt-Ingram Cancer Center, Nashville, USA;7. INCLIVA, Biomedical Research Institute, Hospital Clínico Universitario, University of Valencia, Valencia, Spain;8. University Medical Center Mainz, Mainz, Germany;9. BC Cancer/University of British Columbia, Vancouver, Canada;10. Clinic Favoriten, HPB Center Health Network Vienna and Sigmund Freud University, Medical School, Vienna, Austria;11. University Hospitals Gasthuisbergs, Leuven;12. Katholieke Universiteit Leuven, Leuven, Belgium;13. Department of Oncology, OncoHealth Institute, Fundación Jiménez Díaz University Hospital, Madrid, Spain;14. Department of Medical Oncology, The Christie NHS Foundation, Manchester;15. Division of Cancer Sciences, University of Manchester, Manchester, UK;16. Institut du cancer Paris CARPEM, APHP, Georges Pompidou Hospital, Université Paris Cité, Paris, France;17. Icahn School of Medicine at Mount Sinai, Mount Sinai Liver Cancer Program, New York, USA;18. Institut d’Investigacions Biomèdiques August Pi i Sunyer Hospital Clínic, Universitat de Barcelona, Barcelona;19. Institució Catalana de Recerca i Estudis Avançats, Barcelona, Spain;20. University of Leipzig Medical Center, Comprehensive Cancer Center Central Germany, Leipzig, Germany;21. Vall d’Hebron Hospital Campus, Barcelona, Spain;22. Institute of Oncology, IOB-Quiron, UVic-UCC, Barcelona, Spain;23. American University of Beirut, Beirut, Lebanon;24. Aichi Cancer Center Hospital, Nagoya, Japan;25. Masaryk Memorial Cancer Institute, Faculty of Medicine, Masaryk University, Brno, Czech Republic;26. Instituto Alexander Fleming, Buenos Aires, Argentina;27. Karolinska University Hospital, Karolinska Institutet, Stockholm, Sweden;28. Tampere University Hospital, University of Tampere, Tampere, Finland;29. Henry Ford Cancer Institute, Departments of Oncology and Pharmacology, Wayne State University, Detroit, USA;30. Comprehensive Cancer Center, Medical University of Vienna, Vienna, Austria;31. Instituto Nacional de Cancerologia, Mexico, Mexico;32. Clinica Universidad de Navarra and CIBEREHD, Pamplona, Spain;33. Ulm University Hospital, Ulm, Germany;34. Vall d’Hebron Hospital Campus and Institute of Oncology, IOB-Quiron, UVic-UCC, Barcelona, Spain;35. Hammersmith Hospital, Imperial College London, London, UK |
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| Abstract: | - 1.1. Mammalian major apurinic/apyrimidinic (AP) endonuclease, APEX nuclease (Mr 35.4 kDa) was purified from HeLa cells. A hybrid protein (Mr 36.4 kDa), which was expressed in BW2001 strain cells of E. coli, comprising human APEX nuclease headed by 10 additional amino acids was also purified.
- 2.2. The purified preparations were frequently associated with 31-, 33- and 35-kDa peptides having AP endonuclease activity.
- 3.3. The 33- and 35-kDa peptides were suggested to be formed from the hybrid protein or APEX nuclease during their purification processes by proteolytic cleavage with subtilisin-like protease. The 31-kDa peptide was thought to be produced by chemical cleavage of the aspartyl-prolyl bond of APEX nuclease.
- 4.4. The results support the notion that some of AP endonuclease heterogeneity based on the molecular weight difference are caused by proteolytic (and chemical) cleavage of a species of AP endonucleases during the extraction and purification.
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