Purification and biological characterization of human hepatopoietin A, a polypeptide growth factor for hepatocytes - PubMed (original) (raw)
. 1989 Jun 15;49(12):3314-20.
Affiliations
- PMID: 2524251
Purification and biological characterization of human hepatopoietin A, a polypeptide growth factor for hepatocytes
R Zarnegar et al. Cancer Res. 1989.
Abstract
We have previously reported the presence of a high molecular weight polypeptide growth factor in the plasma of normal human or rat serum which stimulates DNA synthesis in primary cultures of normal rat hepatocytes. We referred to this activity as hepatopoietin A (HPTA) (Michalopoulos, G., Houck, K. A., Dolan, M. L., and Luetteke, N. C. Control of hepatocytes replication by two serum factors. Cancer Res., 44: 4414-4419, 1984; Thaler, J., and Michalopoulos, G. Hepatopoietin A. Partial characterization and trypsin activation of a hepatocyte growth factor. Cancer Res., 45: 2545-2549, 1985). At that time, however, complete purification of this growth factor had not been achieved. In the present report we describe the steps required for complete purification of HPTA from human plasma or rabbit serum. The purification involved sequential ammonium sulfate precipitation, heparin-affinity chromatography, anion-exchange high-performance liquid chromatography (HPLC), and reversed phase HPLC. The final purified product is a heterodimer consisting of a heavy and a light polypeptide chain with molecular weights of 70,000 and 35,000, respectively, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Under nonreducing conditions, however, the purified HPTA migrated as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis corresponding to a molecular weight of 69,000. The mitogenic activity of HPTA was associated with this band when it was eluted from unstained sodium dodecyl sulfate-polyacrylamide gels. Gel filtration HPLC under neutral isotonic conditions indicated that HPTA tends to form aggregates with molecular weights of greater than 300,000. Chromatofocusing indicated that HPTA is an acidic protein with an isoelectric point value of about 5.5. The mitogenic activity of HPTA was sensitive to heat, trypsin, and 2-mercaptoethanol, but relatively resistant to exposure to 1 N acetic acid, 2 M guanidine-HCl, and 0.1% sodium dodecyl sulfate. The stimulation of DNA synthesis induced by HPTA was totally abrogated by transforming growth factor-beta and markedly reduced in the presence of heparin. We present biochemical as well as biological evidence that HPTA is a hepatocyte growth factor distinct from other known polypeptide mitogens such as epidermal growth factor, transforming growth factor-alpha, platelet-derived growth factor, fibroblast growth factor, and thrombin.
Similar articles
- Hepatopoietin A: partial characterization and trypsin activation of a hepatocyte growth factor.
Thaler FJ, Michalopoulos GK. Thaler FJ, et al. Cancer Res. 1985 Jun;45(6):2545-9. Cancer Res. 1985. PMID: 3157446 - Characterization of the effects of human placental HGF on rat hepatocytes.
Hernandez J, Zarnegar R, Michalopoulos GK. Hernandez J, et al. J Cell Physiol. 1992 Jan;150(1):116-21. doi: 10.1002/jcp.1041500116. J Cell Physiol. 1992. PMID: 1370501 - Purification and characterization of a novel transforming growth factor.
Halper J, Moses HL. Halper J, et al. Cancer Res. 1987 Sep 1;47(17):4552-9. Cancer Res. 1987. PMID: 2887280 - Liver regeneration: molecular mechanisms of growth control.
Michalopoulos GK. Michalopoulos GK. FASEB J. 1990 Feb 1;4(2):176-87. FASEB J. 1990. PMID: 2404819 Review. - Liver-specific growth factors.
Fleig WE. Fleig WE. Scand J Gastroenterol Suppl. 1988;151:31-6. doi: 10.3109/00365528809095911. Scand J Gastroenterol Suppl. 1988. PMID: 2976174 Review.
Cited by
- Assessing self-renewal and differentiation in human embryonic stem cell lines.
Cai J, Chen J, Liu Y, Miura T, Luo Y, Loring JF, Freed WJ, Rao MS, Zeng X. Cai J, et al. Stem Cells. 2006 Mar;24(3):516-30. doi: 10.1634/stemcells.2005-0143. Epub 2005 Nov 17. Stem Cells. 2006. PMID: 16293578 Free PMC article. - From chronic liver disorders to hepatocellular carcinoma: Molecular and genetic pathways.
Ierardi E, Rosania R, Zotti M, Giorgio F, Prencipe S, Valle ND, Francesco VD, Panella C. Ierardi E, et al. World J Gastrointest Oncol. 2010 Jun 15;2(6):259-64. doi: 10.4251/wjgo.v2.i6.259. World J Gastrointest Oncol. 2010. PMID: 21160638 Free PMC article. - Selective induction of DNA synthesis in mouse preneoplastic and neoplastic hepatic lesions after exposure to phenobarbital.
Klaunig JE. Klaunig JE. Environ Health Perspect. 1993 Dec;101 Suppl 5(Suppl 5):235-9. doi: 10.1289/ehp.93101s5235. Environ Health Perspect. 1993. PMID: 8013413 Free PMC article. - The Gab1 PH domain is required for localization of Gab1 at sites of cell-cell contact and epithelial morphogenesis downstream from the met receptor tyrosine kinase.
Maroun CR, Holgado-Madruga M, Royal I, Naujokas MA, Fournier TM, Wong AJ, Park M. Maroun CR, et al. Mol Cell Biol. 1999 Mar;19(3):1784-99. doi: 10.1128/MCB.19.3.1784. Mol Cell Biol. 1999. PMID: 10022866 Free PMC article. - Hepatocyte Growth Factor Isoforms in Tissue Repair, Cancer, and Fibrotic Remodeling.
Mungunsukh O, McCart EA, Day RM. Mungunsukh O, et al. Biomedicines. 2014 Nov 5;2(4):301-326. doi: 10.3390/biomedicines2040301. Biomedicines. 2014. PMID: 28548073 Free PMC article. Review.
Publication types
MeSH terms
Substances
LinkOut - more resources
Other Literature Sources