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Purification and physical‐chemical characterization of hepatic stimulator substance
Author(s) -
Labrecque Douglas R.,
Steele Gregory,
Fogerty Steven,
Wilson Michelle,
Barton James
Publication year - 1987
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840070121
Subject(s) - characterization (materials science) , chemistry , medicine , nanotechnology , materials science
Hepatic stimulator substance is a liver growth stimulator derived from the hepatocyte cytosol of weanling or regenerating adult rat livers. The present paper reports the almost 9,000‐fold purification of hepatic stimulator substance with an approximately 100,000‐fold increase in specific growth stimulator activity. Purification steps included heating at 95°C for 15 min, 40% cold ethanol precipitation, passage over Procion Red HE3B, DEAE cellulose and Sephadex G75 columns and gel filtration and reverse‐phase fast protein liquid chromatography techniques. As little as 27 ng per ml of the purest material produced a 2‐fold stimulation in the standard HTC cell activity assay. Further studies indicate that hepatic stimulator substance is a highly negatively charged protein and that disulfide bonds or a complex tertiary structure are not essential to its activity. Hepatic stimulator substance is stable over a wide range of pH's and temperatures. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis with silver stain revealed 1 major band at 12,400 daltons and 1 minor band at 17,500 daltons.