title: Structure of heparin-binding EGF-like growth factor. Multiple forms, primary structure, and glycosylation of the mature protein.
abstract: Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a newly described member of the epidermal growth factor (EGF) family that is mitogenic for BALB/c 3T3 cells, inhibits the binding of 125I-EGF to its receptor, and triggers autophosphorylation of the EGF receptor. HB-EGF was purified from the conditioned medium of U-937 cells using cation exchange, copper affinity, heparin affinity, and two rounds of C4 reversed phase liquid chromatography. The elution profile of the first round of C4 column chromatography contained four growth factor activity peaks with similar specific biological activities. N-terminal and tryptic fragment microsequencing demonstrated that these peaks contained different structural forms of the HB-EGF protein. Some of the differences in the various forms of HB-EGF were found to be due to N-terminal heterogeneity. Microsequencing of tryptic fragments indicated that the mature HB-EGF polypeptide can contain at least 86 of the 208 amino acids predicted by nucleotide sequence to be the HB-EGF precursor molecule. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that the various forms of HB-EGF have apparent molecular masses of 19-23 kDa. Further analysis of the most predominant form of HB-EGF found in U-937 cell conditioned medium indicated that it has a pI of 7.2-7.8 and is O-glycosylated.
authors.affiliation: Department of Surgical Research, Children's Hospital, Boston, Massachusetts.
authors.forename: S
authors.initials: S
authors.lastname: Higashiyama
authors.forename: K
authors.initials: K
authors.lastname: Lau
authors.forename: G E
authors.initials: GE
authors.lastname: Besner
authors.forename: J A
authors.initials: JA
authors.lastname: Abraham
authors.forename: M
authors.initials: M
authors.lastname: Klagsbrun
mesh_terms: D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001681:Biological Assay; D002460:Cell Line; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004261:DNA Replication / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004815:Epidermal Growth Factor / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D006031:Glycosylation; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology; D006493:Heparin / Q000302:isolation & purification* / Q000494:pharmacology; D066257:Heparin-binding EGF-like Growth Factor; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D007525:Isoelectric Focusing; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D010449:Peptide Mapping; D010982:Platelet-Derived Growth Factor / Q000032:analysis / Q000494:pharmacology; D012689:Sequence Homology, Nucleic Acid
chemical_list: D000906:Antibodies; D006133:Growth Substances; C587883:HBEGF protein, human; C587884:Hbegf protein, mouse; D066257:Heparin-binding EGF-like Growth Factor; D036341:Intercellular Signaling Peptides and Proteins; D010446:Peptide Fragments; D010982:Platelet-Derived Growth Factor; D004815:Epidermal Growth Factor; D006493:Heparin