Two forms of murine epidermal growth factor: rapid separation by using reverse-phase HPLC.
- 1 October 1982
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 79 (19), 5753-5757
- https://doi.org/10.1073/pnas.79.19.5753
Abstract
Epidermal growth factor (EGF) was isolated from acid extracts of C57BL6/J mouse submaxillary glands by using hydrophobic chromatography. High yields of EGF in large amounts (10 mg) can be isolated reliably from the acid extract of the glands in less than 4 h. The reverse-phase high performance liquid chromatography techniques used to purify the EGF initially yielded what appeared to be a single homogeneous EGF molecule. However, ion pairing reagents (e.g., heptafluorobutyric acid) altered the chromatographic properties, revealing 2 distinct species: EGF-.alpha. and EGF-.beta.. The apparent MW, isoelectric points and antigenic properties of EGF-.alpha. and EGF-.beta. were identical, and both forms stimulated a mitogenic response in mouse fibroblast 3T3 cells. Analysis of different preparations of purified EGF (commercial and experimental) indicated the presence of EGF-.alpha. and EGF-.beta. in constant proportion. Previous EGF binding studies must have used mixtures of 125I-labeled EGF-.alpha. and 125I-labeled EGF-.beta.. The 2 molecules appear to compete for an identical receptor on the cell surface.This publication has 18 references indexed in Scilit:
- Epidermal growth factor in the submandibular glands of inbred miceJournal of Anatomy, 1981
- USE OF AN OCTADECYLSILICA PURIFICATION METHOD MINIMIZES PROTEOLYSIS DURING ISOLATION OF PORCINE AND RAT RELAXINSEndocrinology, 1980
- Use of hydrophobic interaction methods in the isolation of proteins from endocrine and paraendocrine tissues and cells by high-performance liquid chromatographyJournal of Chromatography A, 1979
- Use of octadecasilyl-silica for the extraction and purification of peptides in biological samples. Application to the identification of circulating metabolites of corticotropin-(1-24)-tetracosapeptide and somatostatin in vivoBiochemical Journal, 1977
- Epidermal growth factor and a new derivative. Rapid isolation procedures and biological and chemical characterization.1972
- The Primary Structure of Epidermal Growth FactorJournal of Biological Chemistry, 1972
- Epidermal growth factor. Physical and chemical properties.1972
- Strategy and tactics in protein chemistryBiochemical Journal, 1970
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Isolation of a Mouse Submaxillary Gland Protein Accelerating Incisor Eruption and Eyelid Opening in the New-born AnimalJournal of Biological Chemistry, 1962