Purification and characterization of a growth factor from guinea pig harderian gland.
A polypeptide growth factor, Harderian gland-derived growth factor (HGDGF), has been purified approximately 43,000-fold from guinea pig Harderian gland by column chromatography on TSK gel DEAE-5PW, blue-Sepharose CL-6B, and Superose 12. The yield was approximately 10%. The Superose 12 fraction was further purified by Aquapore BU-300 reversed-phase chromatography to apparent homogeneity. HGDGF was eluted from TSK gel DEAE-5PW at 0.20-0.35 M NaCl, with a linear gradient of 0.15-0.80 M NaCl and at 2.20 M NaCl from blue-Sepharose CL-6B. The activity of HGDGF toward human embryonic cells (TIG-3) was quantitated, [3H]thymidine incorporation for 48 h being stimulated in a linear and dose-dependent manner. Purified HGDGF has a molecular weight of approximately 13,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and molecular sieve column chromatography. HGDGF is labile to treatment with SH reagents or acetic acid. Both trypsin digestion and boiling decrease the activity of HGDGF. Its pI is 5.1. HGDGF stimulates the multiplication of TIG-3 cells but has no effect on human endothelial cells K2T1 or A2T2 which require fibroblast growth factor for growth. HGDGF appears to differ from other growth factors, suggesting that it is a previously undescribed growth factor.
Yokoyama
Y
Y
Department of Physiological Chemistry and Nutrition, Faculty of Medicine, University of Tokyo, Japan.
Kano
K
K
Kaji
K
K
Seyama
Y
Y
D000818:Animals; D002455:Cell Division; D002460:Cell Line; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002845:Chromatography; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology; D015943:Factor Va / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology; D006168:Guinea Pigs; D006243:Harderian Gland / Q000032:analysis*; D006801:Humans; D007765:Lacrimal Apparatus / Q000032:analysis*; D008970:Molecular Weight; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface / Q000502:physiology
D006133:Growth Substances; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; C061370:factor Va receptor; D015943:Factor Va