Isolation of multiple biologically and chemically diverse species of epidermal growth factor.
We have analyzed several lots of epidermal growth factor (EGF) purified from murine submaxillary glands including "receptor grade" EGF from Collaborative Research and EGF from Boehringer Mannheim Biochemicals. New England Nuclear uses "receptor grade" EGF to produce 125I-labeled EGF. Though these reagents are reported to be homogeneous, we found them to be a mixture of six species. A method was developed to separate this mixture into its component parts. The individual components were chemically characterized and tested for biological potency. N-terminal sequence analysis of the unfractionated EGF-mixture reveals three different sequences starting with residues 1, 2, or 3 of the mature peptide. Each component exhibited different degrees of mitogenic and EGF receptor binding activity indicating that the N-terminal region contributes to the biological response. The species representing the complete EGF peptide is the most active species in all biological assays. A rapid method for purification of homogeneous complete EGF from commercial EGF preparations is described.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004815:Epidermal Growth Factor / Q000031:analogs & derivatives* / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D066246:ErbB Receptors / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D013363:Submandibular Gland / Q000032:analysis*
D000596:Amino Acids; D004815:Epidermal Growth Factor; D066246:ErbB Receptors