Purification and partial characterization of a liver cell proliferation factor called hepatopoietin

Goldberg, Michel

doi:10.1002/jcb.240270310

Cited by 27 publications

(5 citation statements)

References 16 publications

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“…Small intestine and pancreas had the highest specific activity, correlating well with the amount of HPTA present in these tissues. DISCUSSION During the past 10 years several laboratories, including ours, have reported the presence of mitogenic substance(s) in rat serum and human plasma (5,(11)(12)(13)(14)(15), some of which have been purified to high specific activity and partially characterized (8, 9). Recently we purified to homogeneity a large heparin-binding polypeptide hepatocyte growth factor with high molecular weight from human plasma and rat serum that we refer to as HPTA (5,6).…”

Section: Resultsmentioning

confidence: 95%

Tissue distribution of hepatopoietin-A: a heparin-binding polypeptide growth factor for hepatocytes.

Zarnegar

Muga

Rahija

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

125

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Hepatopoietin-A (HPTA) is a heparinbinding polypeptide growth factor which consists of a heavy and a light polypeptide chain with molecular weights of 70,000 and 35,000, respectively. It stimulates DNA synthesis in primary cultures of normal rat hepatocytes in serum-free medium. The complete purification and characterization of HPTA from rabbit serum were reported by us elsewhere. Recently we have determined the amino-terminal amino acid sequence of the rabbit HPTA light chain up to 24 residues and have shown that the sequence is not homologous with other known sequences. [N.B. Human hepatocyte growth factor, recently sequenced by two other groups, is the same molecular species as HPTA.] In the present paper we report the production of a neutralizing polyclonal antiserum raised in chicken against purified rabbit HPTA. This antiserum does not inhibit the mitogenic effect of other potent inducers of hepatocyte DNA synthesis (epidermal growth factor or acidic fibroblast growth factor), nor does it interact with these growth factors in an enzyme-linked immunosorbent assay (ELISA). The antibody recognizes HPTA, as was determined by Western immunoblotting. Since the tissue origin of HPTA is not known, this anti-HPTA antiserum was used to investigate the tissue distribution of HPTA in rabbits by immunohistostaining methods. Acinar cells of the pancreas, neurons of the brain, C cells of the thyroid, ductal cells of the salivary glands, and Brunners glands of the duodenum stained with anti-HPTA antibody. Liver, spleen, thymus, and kidney do not seem to contain appreciable amounts of HPTA. We confirmed these fin gs by extracting and purifying active HPTA from the stained tissues listed above. The anti-HPTA antibody recognizes HPTA purifiled from different tissues, as was determined by ELISA, Western immunoblotting, and immunoneutralization experiments. We have also determined the sequence of the first 24 amino acids of the amino terminus of the light chain of rabbit HPITA and have shown that there is no homology between this sequence and the sequences of any known proteins and polypeptide growth factors (7). Growth factors with properties similar to those of HPTA have also been purified from human plasma or rat platelets by others (8,9).In the present paper we describe the production of a neutralizing polyclonal antibody raised in chicken against purified rabbit HPTA and the establishment of an enzymelinked immunosorbent assay (ELISA) for detection of nanogram quantities of this growth factor. This antiserum was used to investigate the tissue distribution of HPTA in rabbits. MATERIALS AND METHODSExtraction and Purification of HPTA from Different Rabbit Tissues. Rabbit pancreas, thyroid, pituitary, submaxillary gland, parotid gland, intestinal mucosa, spleen, and brain (from 25-50 rabbits) were purchased from Pel-Freez Biologicals. Frozen tissues were homogenized in extraction buffer consisting of phosphate-buffered saline (PBS; NaCI, 8.8 g/l; NaH2PO4 H2O, 0.2 g/l; Na2HPO4, 1.4 g/l), pH 7.4, containing 1 mM phenylm...

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Section: Resultsmentioning

confidence: 95%

Tissue distribution of hepatopoietin-A: a heparin-binding polypeptide growth factor for hepatocytes.

Zarnegar

Muga

Rahija

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

125

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“…In this connection, it is noteworthy that the origin of hepatopoietin, a liver cell proliferation factor which is purified from remnant livers or plasma of partially hepatectomized rats (9)' is proposed to be the Peyer's patches of the gastrointestinal tract (50). A t present, it is unlikely that hepatopoietin and hHGF are the same peptide because several characteristic differences such as molecular weight, stability by treatments with heat and organic solvents and an affinity to the gel (5,30,31), are observed between these two factors.…”

Section: Discussionmentioning

confidence: 99%

Clinical significance of human hepatocyte growth factor in blood from patients with fulminant hepatic failure

et al. 1989

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We have recently found the presence of human hepatocyte growth factor in sera of patients with fulminant hepatic failure and have purified human hepatocyte growth factor from plasma of a patient with fulminant hepatic failure. In this paper, we report the clinical significance of human hepatocyte growth factor in blood from patients with fulminant hepatic failure. The effect of sera or plasma from 17 patients with fulminant hepatic failure on liver cell growth was examined by use of adult rat hepatocytes in primary cultures. Sera or plasma from 16 of the 17 patients with fulminant hepatic failure stimulated DNA synthesis in hepatocytes more effectively than normal human serum. The mean growth-promoting activity for the 17 patients with fulminant hepatic failure was about 16 times higher than that obtained for normal human serum. This growth-promoting activity of the patients' blood was not related to sex, age, clinical outcome of the patients or type of fulminant hepatic failure, but was intimately related to the clinical grade of hepatic coma. Sera or plasma with Grade III and IV coma showed stimulatory activity on DNA synthesis more markedly than sera or plasma from patients with coma of less than Grade II. In the surviving group, this activity decreased as the hepatic coma of patients improved. In fact, this activity of sera from patients at the recovery stage showed no significant increase compared with that of normal human serum. In the group of terminal patients, this activity increased as the coma developed.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…F 150 and Acr-F 4 were tested for trypsin and chymotrypsin sensitivity (13), heat stability, and neuroaminidase sensitivity (14).…”

Section: Analyses Of Physical and Chemical Propertiesmentioning

confidence: 99%

Further steps of hepatic stimulatory substance purification

et al. 1991

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The hepatic stimulatory substance (HSS) extracted from weanling rat livers was purified 381,000-fold using chromatographic techniques including nondissociating polyacrylamide gel electrophoresis (nondenaturing PAGE). The activity of this highly purified HSS, named Acr-F 4 , was assessed in two in vivo models. In 40% hepatectomized rats, it produced a fivefold increase in the proliferative rate normally seen following this partial hepatectomy. In Eck fistula dogs, the level of base increase in hepatocyte renewal was amplified threefold by an infusion of Acr-F 4 (50 ng/kg/day). Acr-F 4 had no influence on the regenerative response of the kidney following a unilateral nephrectomy or of the bowel following a 40% resection of the small bowel. On the basis of these findings, it can be concluded that HSS (Acr-F 4 ) has a high biological activity and is organ specific.

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Purification and partial characterization of a liver cell proliferation factor called hepatopoietin

Cited by 27 publications

References 16 publications

Tissue distribution of hepatopoietin-A: a heparin-binding polypeptide growth factor for hepatocytes.

Tissue distribution of hepatopoietin-A: a heparin-binding polypeptide growth factor for hepatocytes.

Clinical significance of human hepatocyte growth factor in blood from patients with fulminant hepatic failure

Further steps of hepatic stimulatory substance purification

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